The current research focused on the effects of parity, season, and lactation stage on the milk yield of indigenous dairy cows in selected districts of the Kaffa Zone of southern Ethiopia. The districts of Gesha and Chena were purposefully chosen. The study design for the 384 household surveys was a cross-sectional survey with a simple random sample approach as the sampling method. Following the survey, 192 lactating cows were chosen for a monitored investigation to track nighttime and morning milk supply. In addition to key informants and focus group discussions, primary and secondary data were obtained via a semi-structured questionnaire, interview, and field observation. The most favored features functioning as selection criteria were those favored for production, reproduction, physical appearance, physiological function, and temperament, in that order. Evening milk yield (EMY), mornning milk yield (MMY), daily milk yield (DMY), lactation length (LL) and lactation milk yield (LMY) were 0.91 ± 0.033 L, 1.22 ± 0.037 L, 2.125 ± 0.07 L, 6.36 ± 0.116 months, and 427.10 ± 20.678 L, respectively, with significant difference (p < 0.01) amongst districts, parity, and season. However, there was no significant variation between studies in EMY, MMY and DMY. Significantly higher values of production performances were recorded for interaction among districts, parity, and season. The study also revealed that interaction between district (Gesha) and parity (third), which were signficantlly higher for EMY, MME, DMY, LL, and LMY 1.50 L, 2.00 L, 3.50 L, 8.10 months, and 850.50 L, respectively. The same trend also observed for interaction among district (Gesha), season (wet), and parity (third), which were obtained to be highly significant values of EMY, MMY, DMY, LL, and LMY were 1.35 L, 1.75 L, 3.10 L, 7.65 months, and 716.80 L, respectively. The study region has a large population of indigenous dairy cows and produces higher yields than the national average. However, it falls short of the worldwide production benchmarks, and the trait preferences used as a selection criterion were based on the farmers' indigenous knowledge. Measures to enhance production abilities must be incorporated and selection criteria must be modernized. The popThe most prevalent features of favor (preferred) were milk yield and traction power, fertility, body weight, feeding behavior, temperament, color and disease resistance, and hard environmental conditions, which differed according to the production method, cultural tradition, and agro-ecology  . Kaffa Z22.1030' and 36°46' east longitude. It bordered the northern Jimma zone in the north, the Sheka zone in the north-west, Bench-sheko in the south-west, the west Omo zone in the south, and Konta special woreda in the southeast. It is divided into three traditional climatic zones based on height and climatic considerations. The Highland (2500–3000 masl), Midland (1500–2500 masl), and Lowland (500–1500 masl) made up 11.6%, 59.5%, and 28.9 % of the total area, respectively. The yearly temperature ranged from 10.2 to 27.5° Celsius, with rainfall ranging from 100 to 2450 mm. Depending on humidity, the dense rainy season is June–September, the transitional or short rainy season is October, November, and May, and the dry season is December-April [Kaffa zone located between 6° 24' and 8° 13' north latitudes and 352.2The types of study designs were both cross sectional study design for surveys and longitudinal study design for follow up (monitored study) of 192 indigenous dairy cows.2.3Aside from the districts that were specifically chosen, three kebeles from each district were picked using basic random selection approaches . Similarly, 64 farmers were chosen at random from each of the kebeles for the cross-sectional research . For the monitoring project, 32 lactating cows with varying parity numbers and lactation phases were purposefully selected from survey data. The reasons were twofold: first, to avoid data recording stoppage if a cow became dry; and second, to investigate the effects of an equal number of parities and lactation stages on milk production in the evening, morning, and day milk yields using fixed variables.2.4The desired sample size for the study was calculated by the formula given by Thrusfield .Consequently, 95 % confidence interval is required with 5 % margin of error in current study. Hence, sample size (n) is determined by the following formula:Expected population proportion = 50 % (0.5)e = desired absolute precision or the margin of error 5 % (0.05)Zα = is the abscissa of a normal curve that cuts of an area at the tails with 95 % confidence interval and the highest margin of error 5 % having 1.96 in Z table± 0.116 months. Therefore, LL was categorized into three stages: early lactation stage (1–2 months), mid (3–4 months) and late (≥5 months). Moreover, parity number were classified as first, second and third (≥3) parities . Further, for fixed effect of wet and dry seasons of the year, a monitoring study was held for milk yield at evening, morning, and daily milk with intimate follow-up, and lactation milk was calculated by multiplying daily milk yield with lactation length. The amount of milk yield was measured with a volumes calibrated plastic jog.Following the survey, a monitoring study was conducted on 192 lactating indigenous dairy cows. Each district was allocated 96 lactating dairy cow with 32 lactating cows, having different parity and lactation stages in each kebele. For reliable lactation length stratification, survey data was immediately analyzed as soon as it was completed. In this way average lactation length (LL) of indigenous dairy cow in the study area was found 6.36 2.5The types of data sources were both primary and secondary. Primary data were collected from all households, focus group discussions (FGD) and key informants through a well-structured and pretested questionnaire, an interview, and field observation. Key informants encompassed livestock and fishery development office heads, private practitioners, traders, and input suppliers. Group discussions were conducted with top kebele administrators, model farmers, and extension workers having 7–10 members in each kebele. Principled enumerators who graduated from TVET and Dandi boru Animal Science College were selected, trained, and employed with the closest researcher following up for significant and relevant data collection. Secondary data were collected from written documents of the livestock and fishery resource development department and offices, agriculture and natural resource development department and offices, the center of AI, epidemiology reports, veterinary anamnesis records, published journals, articles and books.2.6Collected data on livestock population and land holdings of individual farmers were entered into MS Excel spread sheet Excel, 2010) for data clearance and analysed using stastical analyisis system (SAS) software 9.3 version  for data.(2)IndexWhere: n = the number of trait ranks in concern.Milk production performances such as evening milk yield, morning milk yield, daily milk yield, lactation milk yield, and lactation length were analyzed using general linear model (PROC GLM) procedure of SAS . If ther33.1The cattle population and land holdings per household in the study area are presented in 3.23.2.1The feed type and sources such as grazing land, supplementary and complementary feed are presented in 3.2.2Complete house enclosure (24 %), tethering at nearby home (46.1 %), and extensive layover (29.95 %) were mechanisms by which farmers kept their dairy cows at night in worse climatic condition with no significant variation (p˃0.05) between districts of current study area . Calves According to the key informants, survey responses, and FGD, tethering was to avoid crop damage, to prevent predatory attacks, Proper utilization of natural resources, prevent unwanted breeding and to collect dung for natural fertilizer. The roofs were constructed from corrugated iron and local grass are 5.73 %, and 18.23 %, respectively. Floors made from compact soil and wood products like bamboo tree and timber 19.01 %, 4.95 % with no significant variation (p˃0.05). Cleaning was once (8.1 %), twice a day (6.5 %), and conditionally depending up on accumulation of dirty, mud, and muck accounted (6.25 %) in study area with no significant difference (p˃0.05). Conversily, the majority of farmers (70 %) cleaned dung and urine from dairy cattle house twice a day in morning and evening while (30 %) cleaned it only once per day in morning [3.2.3Watering frequencies and water sources of the current study are presented in Moreover, river 30.3 %, well 24.6 %, supplied pipe/tank 40.5 %, and spring water 4.5 % was reported . Waterin3.2.4Awareness of the farmer about common zoonotic diseases  and the Similarly, Lenco and Seblewongel  reported3.3Trait preferences of dairy cows in the study area were presented in Table  accordinThe study revealed that, high milk yield, high reproduction potential, traction power, high live weight, stress tolerance, feed conversion efficiency, and red coat color were preferred traits in descending order . Afras  reportedBesides milk production, traction power, adaptation, and disease resistance ; growth3.43.4.1Climatic factors are totipotent in the lives of organisms all over the world. Production performances of dairy cows in the study area were presented in The values of lactation milk yield (LMY) obtained in the current study were higher than 225 L of Danakil breed  and 339.3.4.2Stages of maternity (parity) had an effect on the production performance of dairy cows . The pre3.4.3Table  demonstr3.4.4Table  illustra3.4.5(14 h) than day length (10 h) in the cow milking trend in the study area. Besides, the physiological rest of various systems in the dairy cow and complete access to ATP that enhance mammary gland lactogenesis at night are more likely to occur at night than at daytime.The effect of the study on production performances like milk yield at evening, morning, and average (DMY), LMY, and LL presented in 3.4.6Districts, parity, and season interact among themselves for evening milk yield, morning milk yield, daily milk yield, lactation milk yield and lactation length . A simil3.4.7Correlations among different variables were verified in 3.4.83.4.9Milking was once (22.7 %), twice (70.6 %), and three times per day (6.8 %) with highly significant variation between districts . There w3.5Opportunities for indigenous dairy production in the study area were presented in 4Dairy production system of the study area was a mixed crop-livestock production system in which uncharacterized zebu cows were burdened with production responsibilities. Trait preferences which rely on indigenous knowledge of producers, were in favor of the selection criteria. Production performances were below international production performance standards, eventhough they were better than the national average yield. The study also showed that the over all daily milk yield, lactation milk yield, and lactation length were highly influenced by district, parity, seasons and their interactions. Even though indigenous dairy cows were powerhouses of opportunities, welfare abandonment, poor management besides of constraints made them deprived in production aptitudes under ideal standards. Hence, rural dairy cow producers should have to get awareness creation training, and periodic proficiency curriculum for veterinarians and technicians needs to be established. Constraints demanding immediate alleviation should get priority, then identified, and an unconditional reaction has to be given.Animal care and ethical issue were careful evaluated and approved the experiment (1956 ET-12/2020) by MizanTepi University, College of Agriculture and Natural Resources ethics Committee. Directive 2010/63/EU of the European Union guidelines (2010) concerning the treatment and use of animals in research and development purposes were employed.The data that has been used is confidential.No funding was obtained for this study.No additional information is available for this paper.Regasa Begna Roba: Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Project administration, Software, Supervision, Validation, Visualization, Writing – original draft, Writing – review & editing. Yakob Asfaw: Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Resources, Supervision, Validation, Visualization, Writing – original draft, Writing – review & editing. Worku Masho Bedane: Conceptualization, Data curation, Methodology, Supervision, Visualization, Writing – original draft, Writing – review & editing.The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.


Pelvic incidence (PI) minus the lumbar lordosis (LL) angle (PI-LL) correlates with function and disability. It is associated with paravertebral muscle (PVM) degeneration and is a valuable tool for surgical planning of adult degenerative scoliosis (ADS). This study aims to explore the characteristics of PVM in ADS with PI-LL match or mismatch and to identify the risk factors for PI-LL mismatch.A total of 67 patients with ADS were divided into PI-LL match and mismatch groups. The visual analog scale (VAS), symptom duration, and Oswestry disability index (ODI) were used to assess patients’ clinical symptoms and quality of life. The percentage of fat infiltration area (FIA%) of the multifidus muscle at the L1-S1 disc level was measured by using MRI with Image-J software. Sagittal vertical axis, LL, pelvic tilt (PT), PI, sacral slope, and the asymmetric and average degeneration degree of the multifidus were recorded. Logistic regression analysis was done to identify the risk factors for PI-LL mismatch.P < 0.05). There was no statistical difference of asymmetric degeneration degree of the multifidus between the two groups (P > 0.05). In the PI-LL mismatch group, the average degeneration degree of the multifidus, VAS, symptom duration, and ODI were significantly higher than that in the PI-LL match group, respectively (32.22 ± 6.98 vs. 26.28 ± 6.23 (%), 4.33 ± 1.60 vs. 3.52 ± 1.46, 10.81 ± 4.83 vs. 6.58 ± 4.23 (month), 21.06 ± 12.58 vs. 12.97 ± 6.49, P < 0.05). The average degeneration degree of the multifidus muscle was positively correlated with the VAS, symptom duration, and ODI, respectively . Sagittal plane balance, LL, PT, and the average degeneration degree of the multifidus were the risk factors for PI-LL mismatch .In the PI-LL match and mismatch groups, the average FIA% of the multifidus on the convex side was less than that on the concave side (The PVM on the concave side was larger than that on the convex side in ADS irrespective of whether PI-LL matched or not. PI-LL mismatch could aggravate this abnormal change, which is an important cause of pain and disability in ADS. Sagittal plane imbalance, decreased LL, higher PT, and larger average degeneration degree of the multifidus were independent risk factors for PI-LL mismatch. Adult degenerative scoliosis (ADS) is a three-dimensional deformity in skeletally mature individuals, defined as a coronal deviation of greater than 10° . It has The paravertebral muscle (PVM) is closely associated with spinal deformities –7 and hePelvic incidence (PI) minus the lumbar lordosis (LL) angle (PI-LL) can be used to study the relationship between the pelvic and the lumbar curve, and both of them correlate with PVM degeneration , 9, 10. This study aims to reveal the characteristics of the PVM in ADS with PI-LL match or mismatch and identify the risk factors for PI-LL mismatch.This was a retrospective cross-sectional study. Data from patients with ADS who were diagnosed at the Third Hospital of Hebei Medical University during the period from July 2010 to October 2019 were evaluated. The inclusion criteria were as follows: (1) aged ≥50; (2) presence of ADS, defined by a coronal Cobb angle >10°; (3) presence of ADS with no radiculopathy and those who did not receive physical therapy, acupuncture, or brace treatment. The exclusion criteria were as follows: (1) presence of symptomatic spinal stenosis with neurogenic claudication and those who received physical therapy, acupuncture, or brace treatment; (2) a recent history of trauma; (3) a prior diagnosis of scoliosis or other spine deformities; (4) underlying diseases such as diabetes mellitus (DM) and sarcopenia; (5) data integrity. All patients in this study had only symptoms of LBP. The visual analog scale (VAS), symptom duration, and Oswestry disability index (ODI) were used to assess the patients’ clinical symptoms and quality of life.The radiography system used was a 500 mA Siemens DR System  with an automatic exposure control system. The detailed parameters are as follows: the electric current was kept at 500 mA and the voltage at 75 kV on the anteroposterior position, and the current was kept at 500 mA and the voltage at 85 kV on the lateral position.The MRI system was a 1.5 Tesla Imaging System . T1-weighted images (T1WI) and T2-weighted images (T2WI) of sagittal views of the lumbar intervertebral disc were obtained using a spin echo sequence system for T1WI and a fast spin echo sequence system for T2WI. A surface coil was used. The slice width was 4 mm and the interslice gap was 1 mm. The acquisition matrix was 512 × 256. The sequence parameter was repetition time (TR) 482 ms/echo time (TE) 10 ms for T1WI and TR 2,300 ms/TE 99 ms for T2WI.X-ray and MRI were performed for all subjects and the data were recorded in detail.Radiography was a useful tool to evaluate the bony structural parameters of ADS. The radiography consisted of a standing anteroposterior and lateral radiograph of the entire spine. In standard anteroposterior radiography, the lumbar scoliosis Cobb's angle, apical vertebral level, and curve direction of the main/compensatory curve were measured. A distance between the C7 plumb line (C7PL) and the center sacral vertical line (CSVL) of more than 2 cm represented coronal plane imbalance, and if this distance was less than 2 cm, it denoted coronal plane balance . In stanMRI was a valuable tool to assess the changes in the PVM, and T2WI were taken at each L1-SI disc level. The center slice of the multifidus muscle was the primary research subject that included the cross-sectional area (CSA) and the percentage of fat infiltration area (FIA%). The CSA of the multifidus muscle was measured in the following steps. The first step was to set scale pixel/cm, converting pixels into cm measurement units by using the ruler present in the photographic image and converting each image into a grayscale 8-bit image. The second step was to measure the CSA of the multifidus muscle by outlining its region freehand using Image-J software. The measurement of FIA% was based on the above, using a threshold technique. The detailed operation steps were as follows: the value of the threshold was selected by the “default” and “dark background” method automatically and then image/adjust/ threshold/dark background and default/auto. Fat tissue in the 8-bit image was colored red using the threshold technique and the red area was measured as the FIA. FIA% was FIA divided by CSA.χ2 test was used to compare sex between PI-LL match and mismatch. The t-test was used to compare sex, the VAS score, and symptom duration, respectively, between PI-LL match and mismatch. The t-test was used to compare the asymmetric degeneration degree and average degeneration degree of the multifidus muscle between the two groups. The Pearson correlation coefficient was used to assess the correlation between the asymmetric and the average degeneration degree of the multifidus muscle and the VAS score and the symptom duration. Univariate and multivariate logistic regression analyses were used to identify the possible risk factors. For all statistical analysis, the level of significance was set at P < 0.05.Statistical analysis was conducted using IBM SPSS 20.0 software. Non-parametric tests were used to compare age and the ODI between PI-LL match and mismatch. The 2). The subjects were divided into two groups: PI-LL match group (31 cases) and PI-LL mismatch group (36 cases). The mean age of the PI-LL mismatch group was more likely older than that of the PI-LL match group  and the mean body mass index (BMI) was 25.4 ± 1.8 , 22.16 ± 8.70 (%), 25.82 ± 9.70 (%), 25.80 ± 7.10 (%), and 29.89 ± 8.61 (%), and on the concave side, it was 31.87 ± 9.05 (%), 35.18 ± 12.32 (%), 32.49 ± 9.89 (%), 33.32 ± 8.72 (%), and 37.77 ± 10.42 (%) at the L1-2, L2-3, L3-4, L4-5, and L5-S1 levels. The average FIA% of the multifidus muscle on the concave side was larger than that on the convex side at each level , 25.02 ± 9.73 (%), 26.78 ± 9.31 (%), 27.18 ± 7.71 (%), and 31.76 ± 8.64 (%), and on the concave side, it was 36.05 ± 12.14 (%), 38.29 ± 11.85 (%), 35.57 ± 13.19 (%), 36.12 ± 9.60 (%), and 40.95 ± 9.91 (%) at the L1-2, L2-3, L3-4, L4-5, and L5-S1 levels. The average FIA% of the multifidus muscle on the concave side was larger than that on the convex side at each level , and in the PI-LL mismatch group, it was 10.35 ± 5.44 (%), and there was no statistical difference between the two groups , histochemistry, cytology, and pathology to support the findings. In the future, we need to collect more imaging data of patients with ADS through multicenter recruitment and employ more methods of investigation.The PVM on the concave side was larger than on the convex side in ADS irrespective of PI-LL was matched or mismatched. PI-LL mismatch could aggravate this abnormal change, which was an important reason for pain and disability in ADS patients. Sagittal plane imbalance, decreased LL, higher PT, and larger average degeneration degree of the multifidus muscle were independent risk factors for PI-LL mismatch. Strengthening of the PVM may be beneficial in preventing PI-LL mismatch in ADS patients.


As an indicator to evaluate the risk of fracture in diffuse idiopathic skeletal hyperostosis, the maximum number of vertebral bodies’ bone cross-linked with contiguous adjacent vertebrae (max VB) was developed. This study retrospectively investigates the relationship between max VB, bone mineral density (BMD), and bone metabolic markers (BMM).n = 114) with various max VB from the thoracic vertebra to the sacrum, measured using computed tomography scans, were selected to assess femur BMD and BMM. The association of max VB with the total type I procollagen N-terminal propeptide (P1NP), tartrate-resistant acid phosphatase 5b (TRACP-5b), and bone turnover ratio (BTR = TRACP-5b/P1NP) as well as its relationship with femur BMD with P1NP and TRACP-5b, were investigated. Furthermore, the relationship between P1NP and TRACP-5b was investigated.In this cross-sectional study (from April 2010 to January 2022), males (P1NP increased in proportion to max VB and TRACP-5b increased in proportion to P1NP. Moreover, BTR was inversely proportional to max VB. Finally, femur BMD was inversely proportional to P1NP and TRACP-5b.As max VB increased with P1NP—a potential osteogenesis indicator—and BTR was inversely proportional to max VB with compensatory TRACP-5b increase, max VB can be considered as a possible predictor of bone fusion. In diffuse idiopathic skeletal hyperostosis (DISH), a continuous and measurable osteogenesis process involves development from incomplete and pointy bone bridges to more flowing and complete bridges . The forThis retrospective cross-sectional study was conducted in Japan.Between April 2010 and January 2022, a total of 20,357 patients visited the Department of Orthopedic Surgery, Shizuoka City Shimizu Hospital . Of these, 2176 were tested for BMD by dual-energy X-ray absorptiometry . Overall, BMM was evaluated in 312 male patients. Only male patients were selected to eliminate the effects of postmenopausal osteoporosis because DISH occurs more frequently in men. Computed tomography (CT) scans  of the thoracic to lumbar vertebrae and pelvis were obtained from 179 patients. By examining medical records, examination interviews, and phone interviews, we identified 132 patients without an exclusion criterion, i.e., history of hyperparathyroidism and rheumatism, steroid use, and osteoporosis drug treatment. Of 132 cases, we excluded 14 patients with sacroiliac (SI) joint ankylosis, leaving 118 patients without ankylosis. Of these 118 patients, 114  were selected from among male patients only, excluding 4 who had fractures within 1 year of the BMM collection.For the 114 patients, CT scans were obtained from the thoracic vertebra to the pelvis. CT is mainly taken for close examination of lumbar back pain and for determining surgical procedures for degenerative diseases of the spine. In addition, max VB was evaluated from the thoracic vertebra to the sacrum in consultation with three orthopedic surgeons. As per Resnick’s definition, the DISH group included patients with four or more consecutive vertebral cross-links and the non-DISH group included those with fewer than four consecutive vertebral cross-links . Besidesn = 69) and non-DISH (n = 45) groups without adjustment. Furthermore, the outcomes influenced by max VB (n = 114) were investigated. Notably, the outcomes examined were P1NP, TRACP-5b, and BTR, and the confounding factors were age, eGFR, and HbA1c. Furthermore, we examined whether exposure to P1NP and TRACP-5b influenced femur BMD, with age, eGFR, HbA1c, and BMI as the confounding factors. Finally, the influence of P1NP on TRACP-5b was examined with age, eGFR, and HbA1c as confounders.First of all, we investigated the timing of CT, bone mineral density, and bone metabolism markers to investigate the interval between each. Next, the mean and standard deviation were compared with the following items: age, BMI, total serum calcium concentration, serum phosphorus concentration, TRACP-5b, P1NP, eGFR, HbA1c, femur BMD, and BTR between the DISH  and statistical software R-4.0.3 (Index of /src/base/R-4 [r-project.org]). Missing data are 1; eGFR, 4; HbA1c. In addition, comparisons between the DISH and non-DISH groups were conducted for each item using the Mann–Whitney 2, HbA1c 6 ± 0.6%, max VB 5.9 ± 4.7, femur BMD 0.9 ± 0.2 g/cm2, and BMI 19.5 ± 3.1 kg/m2. The number of max VB was as follows: 0 (n = 19), 2 (n = 10), 3 (n = 16), 4 (n = 14), 5 (n = 3), 6 (n = 8), 7 (n = 6), 8 (n = 6), 9 (n = 5), 10 (n = 2), 11 (n = 4), 12 (n = 8), 13 (n = 5), 14 (n = 4), 15 (n = 2), and 17 (n = 2). The number of cases per max VB was evenly divided. Age, eGFR, max VB, P1NP, femur BMD and BTR remarkably differed between the DISH and non-DISH groups in an unadjusted comparison (Table The mean and standard deviation of the timing of CT and BMD were 57 ± 82 days (min 0 and max 364), CT and BMM were 49 ± 79 days (min 0 and max 338), and BMD and BMM were 23 ± 49 days (min 0 and max 287). Overall, the subjects’ mean age was 77 ± 9 years, P1NP 53 ± 28 ng/ml, TRACP-5b 433 ± 177 mU/dl, BTR 9 ± 3.5, eGFR 60 ± 18 ml/min/1.73 mCorrelation coefficient = 0.05, P = 0.003; Fig. The results of this analysis suggested that max VB may have influenced the values of P1NP and femur BMD. Therefore, we first investigated the relationship between max VB and P1NP with age, eGFR, and HbA1c as confounding factors. We found that only P1NP increased in proportion to max VB , are high in AS. Of note, higher CTX signifies lower bone density values. Both AS and DISH are similar in terms of the formation of bone bridges in the spine when bone resorption markers increase, but they differ in bone density and P1NP –25. The To the best of our knowledge, no paper has yet examined the effect of the number of bone cross-links on BMM. Moreover, as mentioned above, studies about BMM comparing DISH and non-DISH have not eliminated confounding factors , 13, 14.In the present study, BMD showed a negative correlation with both bone formation and resorption markers, suggesting that high BMD and bone formation markers are incompatible. An interesting paper examined the correlation between P1NP levels and hip bone density in teriparatide-treated patients, reporting that approximately 60% of patients presented with less hip BMD, which was independent of the changes in P1NP levels . This faHigh bone density is reported to be favorable for bone healing , 33. In This study has some limitations that are worth acknowledging. First, this was a retrospective study, with a small sample size. Second, this study was analyzed using P1NP, which reacts at the early stage of bone formation, rather than osteocalcin or bone alkaline phosphatase, which is a marker of the final stage of bone formation, because the study was conducted within insurance. Finally, that bone cross-link formation in DISH is not only affected by factors owing to bone metabolism, but also by degeneration. In the future, it will be necessary to investigate bone healing after surgery for chronic spinal diseases and fractures with the degree of bone cross-linking in a prospective study.This study demonstrates that max VB increased in proportion to P1NP, which was considered an indicator of bone formation. Furthermore, there was a compensatory rise in the levels of bone resorption and bone rotation increased, whereas BTR was inversely proportional to max VB, suggesting that it could be a predictor of bone healing. Hence, measuring max VB before the spinal fusion surgery is beneficial to easily confirm the possibility of bone fusion.


Bifidobacterium animalis subsp. lactis BB-12, Chr. Hansen, Denmark) and prebiotics (inulin), which had adequate organoleptic characteristics. Moreover, adequate physicochemical, microbiological, and sensory characteristics during its shelf life were expected. A pasteurized skim milk curd flavored with a guava pulp was stabilized with gelatin to formulate this product. As sweeteners, iso maltol, erythritol, and Luo Han Guo extract from monk fruit (Siraitia Grosvenorii) were added. The prebiotic used was inulin, and the probiotic . The product was kept refrigerated (4 °C) during the shelf life of 28 days. For the organoleptic analysis (100 consumers), the evaluations performed were: (1) overall liking (OL), (2) CATA  testing 19 attributes, and (3) purchase intention was evaluated. Results were analyzed with FIZZ Software Biosystèmes. During shelf life, (1) physicochemical, microbiological, and sensory tests were performed. The product was evaluated as “liked much'' (7.16 out of 9); it was described as a creamy (71 %) natural product (73 %) with a fruity odor (57 %). It could be suitable for marketing because 82 % of the consumers would buy it. The product's probiotic character (over 1 × 106) was established through a microbiological count. On day one, the CFU was found to be 4.15 × 108, and after 28 days, 1.98 × 108 CFU of viable Bifidobacterium animalis subsp. lactis BB-12, leading us to establish its probiotic characteristics. The shelf life was estimated at 21 days.The consumption of functional dairy products continues to rise due to consumer needs. This study aimed to develop a dairy guava functional symbiotic petit cheese product that included probiotics  are good sugar replacers in petite Suisse products.•Inulin can proved adequate sensory characteristicas in petit Suisse products.•Of the 100 consumers recruited for sensory evaluation, 82 % would buy the product. Extensive clinical studies have demonstrated the favorable impact of this bacterium on health. These benefits include improvement of the following matters:(1)Gastrointestinal health and immune function ;(2)An increase of the presence of probiotics could contribute to the dental well-being ;(3)Steady improvement of glucose metabolism and insulin sensitivity during and after pregnancy ;(4)Efficacy in alleviating colic, resulting in reduced infant crying and irritability ;(5)Increased survival of BB-12 through the gastrointestinal tract Prebiotics can be defined as a substrate that can stimulate the growth of host microorganisms, thus conferring health benefits ,13. PrebOn the other hand, beneficial bacteria belonging to the genera Lactobacillus and Bifidobacterium grow in the gastrointestinal tract . Further1)Relief from constipation,2)Increased mineral absorption,3)Intestinal pH reduction,4)Intestinal bacterial balance restoration, and modulation of lipid metabolism,5)Anticarcinogenic effects6)Impacts on the immune systemSeveral benefits have been associated with prebiotics :1)ReliefIn particular, inulin compounds are integral to a non-digestible polysaccharide called fructans. Therefore, inulin is a functional component , which cAfter COVID, there is a worldwide trend for sugar reduction, with natural sweeteners; which means replacing refined sugar with alternative sweeteners due to the potential health benefits offered by other ingredients ,18. RefiGuava was selected in this study due to its delightful flavor profile. In addition, it holds the distinction of being the most extensively cultivated fruit in Aguascalientes, Mexico. Its widespread availability throughout the year and its nutritional and health-promoting attributes make it one of the most consumed fruits. Numerous studies have identified bioactive compounds like phenolic acids (caffeic acid), flavonoids (quercetin and kaempferol), and carotenoids (beta-carotene and lycopene) with guava. These compounds play a pivotal role in preventing cerebrovascular, cardiovascular, and cancer-related issues, thus enhancing the functional impact of the product ,24,and25Bifidobacterium animalis subsp. lactis BB-12, Chr. Hansen, Denmark) and prebiotics (inulin) integrated in the dairy matrix. The resulting product was monitored throughout its shelf life in its physicochemical, microbiological and sensory changes. The product development was conducted at the food technology laboratory of the Universidad Panamericana Aguascalientes, Mexico.Therefore, the objective of this study aimed to develop a functional guava symbiotic petit cheese product; the food system developed included probiotics . Then it was mixed in the thermomix  at speed 6 for 3 min. A seed separation was performed through a stainless steel strainer, according to procedure established in Siraitia grosvenorii), iso maltol, salt, and citric acid were added to the pulp mixing all in the thermomix  at speed 3 for 1 min . The pulp was cooled to 4 °C.2.1.1.2As observed in 2.1.1.3Bifidobacterium animalis subsp. lactis BB-12 was incorporated in 2.09 %; as recommended by the supplier . The latter for obtaining a bacterial load higher than 1 × 106 CFU/g in the final product. For which, the lyophilized Bifidobacterium animalis subsp. lactis BB-12, culture (5 g) was diluted in (40 mL) of pasteurized skimmed milk and was incubated at 37 °C for 24 h.The prebiotic, inulin , was added in a 0.32 %. On the other hand, 2.1.1.4The gelatin , which had a 290 Bloom, was hydrated by diluting it with water in a 1:2 ratio. The final concentration of this mixture was 0.28 % of the total concentration of the formulation.2.1.2In order to prepare the Petite Suisse cheese, all the ingredients  were mix2.1.3The samples were vacuum-sealed in bags and refrigerated at temperatures ranging from 0 to 4 °C for storage. Product's shelf life was evaluated.2.2The following analysis were performed by triplicate.2.2.1Humidity was determined, by triplicate, calculating weight loss due to water evaporation using the methodology described in NOM-243-SSA1-2010) . The cur2.2.2Color was measured, by triplicate, using the colorimeter . The measurement was performed directly on the packaging (transparent bag), placing the colorimeter on the surface. The results obtained were the L*a*b* coordinates of the CIELAB system and captured through a smartphone.2.2.3pH, which was performed in triplicate, was measured using the methodology described in NOM-F-317-S-1978 [7-S-1978  with a p2.2.4Total soluble solids, which were performed using a refractometer . For the analysis, a mixture drop was put into the equipment for measurement. Total soluble solids were determined in the pulp by triplicate.2.2.5Titratable acidity was performed by triplicate. It was carried out with a NaOH solution at 0.1 N. . The indicator was phenolphthalein 1 %, which was diluted in 70 % ethanol , using the methodology described in NOM-243-SSA1-2010 . The mil2.2.6Syneresis was evaluated visually in the Petite Suisse on the vacuum-sealed bag over time. A subjective measure was employed to characterize the product, and the parameters were used according to a scale elaborated in the Universidad Panamericana. The scale used four parameters to describe the product: not present, slightly present, present, and excessively present.2.3For the organoleptic analysis, 100 consumers ranging from 18 to 23 years on average, who were students from Universidad Panamericana Aguascalientes, participated in this study. All consumers agreed to participate in this research and signed an Informed Consent Form. They answered a sensory questionnaire that evaluated the sensory characteristics of the Petite Suisse.Samples, which were previously coded with a 3-digit identifier, were served in 1-ounce plastic cups that contained 30 g of the product and were served at 6 °C.The questionnaire was divided into three sections: (1) liking evaluation, (2) CATA Test and (3) Purchase intention.2.3.1First, the consumers were asked to grade the acceptance of the product on a 9-point hedonic scale.2.3.2Secondly, the CATA test  that had nineteen attributes was applied to consumers. Consumers were instructed to check the attributes that they found present in the product. The attributes evaluated were: The attributes studied were: pleasant view, natural color, artificial color, pleasant smell, fruity flavor, no odor, tasteless flavor, artificial flavor, natural flavor, fruity flavor, milky flavor, lack of flavor, creamy, sweet, acidic, firm texture, fluid texture, good aftertaste and lumpy ,29.2.3.3Finally, consumers answered whether they would buy the food product or not.2.4The shelf life study was conducted in real-time for 28 days, taking samples on days 1, 7, 14, 21, and 28. The following physicochemical parameters were determined (triplicate): a) humidity, b) color, c) tiratable acidity, and d) syneresis. The methodology of these tests is explained in section E. coli, and total coliforms can be observed. On the other hand, in order to evaluate the functional effect of the product, a viable bacterial count was performed on Bifidobacterium animalis subsp. lactis BB-12 through microbiological analysis of lactic acid bacteria. The methodology is explained in section The methodology for microbiological analysis is explained in section The sensory test was performed using the methodology of RATA . This sensory test was carried out to evaluate the sensory characteristics. The product was sensory evaluated on days 1, 7, and 21.2.4.12 and a temperature of 120 °C. Once the pressure and temperature were achieved, they were maintained for 15 min. Next, 10 g of the sample was weighed and placed in a bag , and 90 mL of sterile water was added in order to obtain a dilution of 10 −1. For which, a sample volume with 1 ml of the dilution 10−1 was used. The microbiological analyses were performed in a laminar flow hood . The growth of total aerobic bacteria in was obtained in 3 M™ Aerobic Count Plates Petrifilm™. For the E. coli, 3 M™ E. coli Petrifilm™, were used. And for the coliform count, 3 M™ Coliforms 6404 Petrifilm™, were used. The incubation  took place under different parameters according to the type of microorganism. For aerobic total bacteria, the incubation parameters were 48 h at 35 °C ± 1. The parameters for coliforms were 24 h at 37 °C ± 1. Finally, for E. coli 48 h at 37 °C ± 1. The counting of viable cells and interpretation of results were carried out as suggested by the supplier of the 3 M México plate [The material was prepared for the microbiological analysis as follows: To 250 mL flasks , 90 mL of distilled water was added. Test tubes were prepared with 9 mL of distilled water, and the micropipette was sterilized in a vertical autoclave . The autoclave was heated until reaching a pressure of 1.5 kg/cmco plate ,31. The 2.4.2−2 dilution, 1 mL was transferred to a test tube containing 9 mL of sterile water. This blend was further mixed using a vortex mixer , and the process was repeated successively to obtain dilutions up to of 10–6. The microbiological analyses were performed in a laminar flow hood . For the plating methodology, the incubation took place at 48 h at 37 °C ± 1, and the counting and interpretation of the results were carried out as established by the supplier of the 3 M México plates [A microbiological analysis of lactic acid bacteria was performed in order to evaluate the functional effect of Petit Suisse cheese. As in the other microbiological analysis, the material was previously sterilized. The sample was prepared using a peristaltic homogenizer . In addition, 1 ml of the dilution was inoculated on the 3 M™ Petrifilm™ Lactic Acid Bacteria. Later on in order to create the 10o plates . This mi2.4.3A sensory analysis was performed to determine the attributes that could describe the guava Petit Suisse cheese shelf life changes over time as shown in The samples (30 g) were presented in a 1-ounce cup coded with a 3-digit number. The judges were invited to participate during the morning on days 1, 7, and 21 of the product's shelf life.Judges were asked to try the product and answer a sensory questionnaire, which consisted of a Rate All that Apply (RATA) test, which first analyzed if an attribute was present and if it was to be rated accordingly. The following attributes were monitored: natural color, fruity odor, milky flavor, rancid flavor, bitter flavor, and firm texture.Semi-trained judges were asked to evaluate how much they liked the product using an unstructured intensity scale, represented by a line of 11 cm that represented their beginning and end with the minimum and maximum degrees of acceptance.2.52.5.1The determinations are presented as averages with standard deviations, and the analyses were conducted in triplicate.2.5.22.5.2.1This study was performed after the product was made. Data was analyzed by the system FIZZ .2.5.2.2O): The data obtained from the consumer's level of satisfaction are typically distributedNull hypothesis (H1): The data obtained from the level of consumer satisfaction are not normally distributed.Alternative Hypothesis  that consumers like guava Petit Suisse cheese equally.1): Consumers do not like guava Petit Suisse cheese similarly.Alternative hypothesis . Through this system, a principal component analysis (PCA) was performed. PCA is a statistical test that aims to represent the variation in the data set. Visual analysis is usually presented in two or three dimensions; in this study, it is presented in two dimensions; each dimension shows the percentage of explainability that each axis has . Therefo33.1After the triplicate determinations, the pasteurized and skimmed milk had a pH of 6.7 and an acidity of 17°D. These values aligned with the characteristics of fresh and sweet milk with a pH between 6.6 and 6.7 and a titratable acidity of 15–17°D . These vThe guava pulp yielded a pH of 3.1, lower than that reported by Medina & Pagano , recordeInulin possibly contributed to improving the texture of the product due to its techno-functional properties, which have been reported to modify texture, which makes it a promising ingredient for dairy products. Therefore, Inulin not only helps with texture but also contributes to its prebiotic character ,16.While Villegas de Gante  indicate3.2The attributes in On the 9-point hedonic scale, an average of 7.16 was obtained, corresponding to “I like it a lot,” with a standard deviation of 1.62.The Kolmogorov-Smirnov statistical test yielded a value of 0.1283, at a significance level of 1 %. For which this value was compared using this value to the critical value of 0.1031 from the tables at the same significance level, the calculated value surpasses the critical value, leading us to reject the null hypothesis . Since d0 and concluding that substantial evidence suggests that consumers do not equally favor the guava Petit Suisse cheese. This assertion is reinforced by 82 % of consumers' intention to buy the product from supermarkets.The calculated chi-square value with a significance of 1 % was 37.54, surpassing the critical of 24.054 obtained from tables at the same significance level. The comparison indicates that the calculated chi-value is higher than the critical value, prompting the rejection of H3.3The shelf life was determined based on physicochemical, microbiological, and sensory analyses. The results of the tests conducted in real-time for 28 days are presented below.3.3.1The guava Petit Suisse cheese was packed in transparent high-vacuum bags and refrigerated at temperatures ranging from 0 to 4 °C. Throughout the storage period, the humidity was maintained within a range of 68.05–71.47 %. This slight variation in humidity can be attributed to the release of moisture within the container, which condenses due to the temperature drop .Evidence of syneresis appeared slightly (Syneresis slightly present) on day 14, as shown in The acidity in the Petit Suisse product changes significantly during storage, exhibiting discernible variations over the storage period. The statistical analysis of acidity during storage shows notable alterations, starting at 0.65 ± 0.011 and culminating at 0.88 ± 0.011 by the end of the product's shelf life.In order to determine the color change over time in guava Petit Suisse cheese, the CIELAB color space, widely used for evaluating food color , was emp3.3.2E. coli. It can be observed from the table that there was no growth of mesophilic aerobic bacteria during the 21 days. Total aerobic count reflects the sanitary quality of food, handling practices, and the hygienic conditions of raw materials [aterials . The groE. coli was absent, in compliance with the maximum permissible limit of 100 cfu/g established by NOM-243-SSA1-2010 [The behavior of total coliforms mirrored that of aerobic mesophilic bacteria. No growth was observed during the initial 21 days, with a count of 10 cfu/g on day 28 . These bSA1-2010 .3.3.36 CFU/g to exert clinical and therapeutic effects on consumers [8 CFU/g on day seven. This count slightly decreased over the days, changing to 1.98 × 108 CFU/g to 8.29 log10 CFU/g on day 28 . Packaging the Petit Suisse cheese in transparent vacuum bags could affect carotenoid stability, as light can lead to their breakdown and the formation of colorless, low molecular weight compounds [The natural color perception decreased until day 21, possibly due to the degradation of the natural pigments in the guava. A study by Ventosa et al.  identifiompounds . A pH deompounds .The fruity odor decreased over 21 days, which could be attributed to the loss of low molecular weight and highly volatile aroma compounds generated by lipid oxidation .The rancid taste increased considerably on day 21. The changes could be due to the biochemical process called lipolysis, a biochemical process transforming triglycerides into partial glycerides and free fatty acids . Free faThe bitter taste increased during the 21 days, particularly in the third week. This phenomenon could be linked to the proteolysis generated in fresh cheeses characterized by high humidity and low pH, resulting in an excessively soft texture and bitter taste .On day 21, the milk flavor decreased, likely due to lipolysis and proteolysis reactions generating alternative flavors and aromas, altering the original milk flavor. Other factors, such as environmental conditions, microorganisms, or compound oxidation, could also influence milk flavor .The texture changed over the three-week period, potentially attributed to enzymatic activity attacking the protein matrix, altering sensory properties such as color, elasticity, and cheese texture ,59.4Bifidobacterium animalis subsp. lactis BB-12, Chr. Hansen, Denmark) and prebiotics (Inulin) in a dairy matrix. The product underwent a comprehensive organoleptic evaluation, revealing noteworthy results. Participants rated the product on a 9-point scale, yielding an average of 7.16, corresponding to a strong preference (“I like it a lot”), with a standard deviation 1.62.A guava functional symbiotic petit cheese product was successfully developed, incorporating probiotics  used Chat GPT for translation purposes. The translation was from Spanish to English to improve the quality of the research paper. Later, the authors used Grammarly to check the document's style. After using these tools/services, the author(s) reviewed and edited the content as needed. Therefore, we take(s) full responsibility for the publication's content.The raw data required to reproduce the above findings are available on request to the corresponding author.Victor Iván Morales-Cortés: Formal analysis, Data curation, Conceptualization. Julieta Domínguez-Soberanes: Writing – review & editing, Writing – original draft, Visualization, Validation, Supervision, Software, Resources, Project administration, Investigation, Funding acquisition, Formal analysis, Data curation, Conceptualization. Linda Carolina Hernández-Lozano: Visualization, Validation, Supervision, Project administration, Methodology, Investigation. Carmen C. Licon: Writing – original draft, Methodology, Investigation, Conceptualization. Antonio Estevez-Rioja A: Methodology, Data curation. Mayeli Peralta-Contreras: Methodology, Data curation.All the authors participated in the development and design of the work and we believe that the manuscript represents valid work, carefully read and fully approved. Moreover, we would like to indicate that there is no bias.So we would like to state a Declaration of interest: None


Carica papaya L.) fruit is a rare phenomenon that may be associated with respiratory metabolism. This study thus aimed to investigate the impacts of storage temperatures (1 and 6 °C) on the respiratory metabolism of postharvest papaya and its impact on CI development. Results demonstrated that 1 °C storage reduced the activities of hexokinase (HK), phosphofructokinase (PFK), pyruvate kinase (PK), citrate synthase (CS), and α-ketoglutarate dehydrogenase (α-KGDH) and regulated the expression of corresponding enzymes in the Embden–Meyerhof–Parnas (EMP) pathway and tricarboxylic acid (TCA) cycle compared with 6 °C storage, resulting in a lower respiration rate of the EMP-TCA pathway and mitigating the development of CI. Meanwhile, lower contents of nicotinamide adenine dinucleotide (hydrogen) (NAD(H)) were observed in papaya fruit stored at 1 °C. Notably, papaya fruit stored at 1 °C maintained higher activity and transcriptional levels of SDH and IDH during the whole storage period. These findings suggest that 1 °C storage reduced the respiration rate of the EMP-TCA pathway by reducing the expression level and activity of related enzymes, which is conducive to the reduction of respiration substrate consumption and finally alleviating the occurrence of CI.Postharvest abnormal chilling injury (CI) behavior in papaya ( Carica papaya L.) originates from Central and South America but is cultivated in all tropical regions [Papaya  in the PPP pathway [Papaya is a climacteric fruit whose respiration rate increases significantly after harvest, and its respiration rate is affected by storage temperature ,10. Resp pathway . However2O2) resulted in increased consumption of metabolic substrates, leading to accelerated damage to the pulp structure and shortened shelf life [The EMP-TCA pathway is the primary respiratory pathway responsible for the utilization of substances to generate energy and other compounds . It has elf life . Tan et elf life  reported2S treatment effectively attenuated the progression of the CI by suppressing the activities of HK, PFK, PK, phosphor-glucose isomerase (PGI), and the levels of glucose and pyruvate, among other components of the EMP-TCA pathway [Respiratory metabolism is a complex physiological process regulated by the enzymatic activities of multiple respiratory pathways . Hexokin pathway . Moreove pathway . Therefore, the present study aimed to investigate the variations in the EMP-TCA pathway in papaya fruit subjected to 1 and 6 °C during storage. Specifically, we analyzed the levels of nicotinamide adenine dinucleotide (NAD) and reduced nicotinamide adenine dinucleotide (NADH), as well as the activities of metabolism-related enzymes and gene expressions involved in the EMP-TCA pathway. The CI index serves as a critical indicator of the severity of CI in papaya fruit . In papa2 kg−1 min−1. In contrast, at 1 °C, the total respiration rate showed a gradual increase, but it was lower than that at 6 °C during the whole storage period , EMP pathway and CI (r = 0.81), and TCA cycle and CI (r = 0.86) (r = 0.61), PFK (r = 0.76), and PK (r = 0.65) were positively correlated with the EMP pathway. α-KGDH (r = 0.84) and IDH (r = 0.59) had a positive correlation with the TCA cycle, but SDH (r = −0.77) had a negative correlation, and CS (r = −0.05) was uncorrelated with the TCA cycle (r = 0.46), CS (r = 0.46), IDH (r = 0.38), and SDH (r = 0.48) were in positive correlation with regulate their own genes, PK (r = −0.43) had a negative correlation with regulate their own genes, but HK (r = 0.06) and α-KGDH (r = 0.14) were uncorrelated with regulate their own genes  and NADH  were uncorrelated with EMP and TCA, but NAD and NADH had a positive correlation with CS , SDH , CpHK , CpPK , and Cpα-KGDH   . ResultsCA cycle . Also, rwn genes . Further = 0.76) .Respiration is essential in the postharvest physiological process of horticultural crops . PostharNAD, acting as an electron acceptor, plays a role in accepting electrons to form NADH in the EMP pathway and TCA cycle ,35. ConsCpHK, CpPFK, and CpPK encoded the above enzyme, which regulated these enzyme functions generated. These genes also regulate the activity of corresponding enzymes in peony [r = 0.61) and PK (r = 0.65)  . Moreoveg et al.  found thg et al. .CpCS, CpIDH, Cpα-KGDH, and CpSDH. Studies have shown that regulating the gene of IDH or α-KGDH in peony [r = 0.84) and IDH activity (r = 0.59) were positively correlated with the respiration rate of the TCA cycle. Additionally, compared to 6 °C storage, 1 °C storage also inhibited the expression level of Cpα-KGDH. These findings collectively suggest that 1 °C storage can reduce the respiration rate of the TCA cycle by suppressing the activities of CS and α-KGDH but promoting IDH activity. Moreover, the SDH activity is a vital indicator to estimate the efficiency of the TCA cycle [CS, IDH, α-KGDH, and SDH are key enzymes in the TCA cycle . The firin peony , the genin peony , and thein peony  can affeCA cycle ,44. NotaCA cycle . PreviouCarica papaya L. cv. Daqing) fruits with a vibrant green color and approximately 10% yellow-green coloration were carefully handpicked from a papaya garden located in the Qiongshan district of Hainan Province, China. The papaya was transported to the laboratory within a 2 h timeframe. For the experiment, only fruit with consistent size, shape, and absence of any flaws or illness indications was chosen.Papaya , including two fruits in each bag. Subsequently, the papaya underwent precooling overnight at 13 ± 1 °C before being transferred to refrigerators set at 1 °C and 6 °C, respectively, for a maximum duration of 48 days. Throughout the storage period, the relative humidity was maintained between 85% and 90%. At intervals of 11 days during storage, twelve fruits were sampled from each group. The fruit peels were rapidly frozen using liquid nitrogen, with a portion of the peel stored in a −40 °C refrigerator for subsequent determination of physiological indices and enzyme activity, and another portion of the peel was stored in a −80 °C refrigerator for the evaluation of gene expression, Additionally, ten fruits per replicate from each group were selected for assessing the CI index. A total of 60 fruits were randomly chosen from each treatment group to participate in the trials, which were performed in triplicate.Initially, the fruit was sterilized for two minutes with a 1 g LThe main symptoms of papaya fruit chilling injury are pitting of the skin, scald, water soaking of the flesh, increased susceptibility to decay, etc. . Among tg. The total, EMP pathway, and TCA cycle respiration rates were reported in terms of μmol O2 kg−1 min−1.The total respiration rate, Embden–Meyerhof–Parnas (EMP) pathway, and tricarboxylic acid (TCA) cycle were determined using a liquid-phase oxygen measurement system  following the methodology described by Tao et al. . To perfg. The obtained result was expressed as mmol kg−1.The determination of NAD(H) contents was performed following a modified method based on Gibon et al.  and Mham2·6H2O, 1 mM EDTA, 5 mM DTT, and 1% (v/v) Triton X-100. After centrifuging the homogenate at 13,000× g for 20 min at 4 °C, the supernatant was collected and used to make the crude enzyme extract.The determination of hexokinase (HK) activity was carried out with certain modifications based on the method described by Zhang et al.  and Wies2·6H2O, 100 μL of 8 mM NADP-Na2, 100 μL of 20 mM ATP, 100 μL of 50 mM glucose, and 50 μL of 40 U mL−1 glucose-6-phosphate dehydrogenase. The absorbance at 340 nm was measured using a UV spectrophotometer . The HK activity was expressed as U kg−1 protein.For the HK assay, the supernatant (0.2 mL) was thoroughly mixed with 650 μL of Tris HCl (20 mM) containing 1.5 mM MgCl−1 protein.Using assay kits purchased from Suzhou Keming Biotechnology Co., Ltd., Suzhou, China, the activities of PFK and PK were determined. The PFK and PK activities were quantified and expressed as U kgg for 15 min at 4 °C. The resulting supernatant (50 μL) was utilized for measuring the activities of CS and α-KGDH using specific ELISA kits obtained from Jiangsu Meibiao Biotechnology Co., Ltd., Yancheng, China. The activity of CS and α-KGDH was expressed as U kg−1 protein.To extract CS and α-KGDH, we mashed and homogenized 0.4 g of papaya exocarp in 1 mL of 0.01 M phosphate buffer (pH 7.4). Following that, the homogenate was centrifuged at 4500× −1 bovine serum albumin, 0.5% (w/v) polyvinylpyrrolidone, and 0.1% (w/v) cysteine. The homogenate was immediately filtered through a four-layer muslin cloth after 15 min of extraction and then centrifuged at 4500× g for 15 min at 4 °C. A second centrifugation was performed on the resultant supernatant at 4 °C and 16,000× g for 30 min. The resulting precipitate was resuspended in a buffer containing 10 mM Tris HCl (pH 7.5), 0.25 M sucrose, 0.3 M mannitol, and 1 mM EDTA. The mixture was stored at 4 °C for activity determinations of IDH and SDH.Extraction of crude mitochondria from papaya exocarp was carried out following the method by Yang et al. , with a 2·6H2O, 1 mM NAD, and 10 mM DL-isocitric acid trisodium salt. The UV spectrophotometer was used to measure the change in absorbance at 340 nm. The IDH activity was expressed as U kg−1 protein.The activity of IDH was determined using the improved method described by Keshavarzian et al. . The 0.5−1 protein.The modified approach outlined by Yang et al.  was usedCpHK (GenBank Accession No. XM_022039184.1), CpPFK (XM_022052063.1), CpPK (XM_022052945.1), CpCS (XM_022031994.1), CpIDH (XM_022036344.1), CpSDH (XM_022051748.1), Cpα-KGDH (XM_022032975.1), and CpActin (FJ696416) in papaya fruit were obtained from NCBI. Primer design was conducted using Primer 6.0 software, and the details of all primers used are provided in −ΔΔCT method reported by Livak and Schmittgen [Using real-time quantitative PCR, the expression levels of genes encoding key enzymes for the TCA cycle and EMP pathway were identified. The gene sequences of K XM_0220063.1, Cpt-test was performed using SPSS 22.0  to analyze the data. Correlations between indicators were analyzed by Pearson’s correlation test. Graphs were generated using Origin 2022b software . p < 0.05 was regarded as statistically significant, whereas p < 0.01 was regarded as very significant, indicating extremely significant differences between groups.With three replicates for each treatment, randomized designs were used across all trials. An independent sample CpHK, CpPFK, CpPK, CpCS, and Cpα-KGDH, indicating that CI alleviation may be closely related to respiration suppression of the EMP-TCA pathway by the regulation of related enzymes and genes. The lower respiration rate of the EMP-TCA pathway contributed to the mitigation of substance consumption. Higher SDH activity was observed in papaya fruit stored at 1 °C, which ensures a sufficient supply of energy. Moreover, the lower NAD content in 1 °C-stored papaya fruit suggests that a portion of NAD is potentially utilized for the biosynthesis of NADPH in the PPP pathway. Consequently, postharvest papaya storage at 1 °C resulted in higher cold resistance compared to storage at 6 °C. The hypothetical model of this research is shown in Storage of 1 °C reduced the activities of HK, PFK, PK, CS, and α-KGDH and regulated the expression of


The temperature decreased towards the distal ROIs compared to proximal ROIs. The average temperatures of the same ROIs were significantly higher for men than women in all regions (p < 0.001). Across all regions, except the dorsal hand region, average temperatures tended to increase with age, particularly in individuals in their 30s and older (p < 0.001). (4) Conclusions: these data could be used as DITI reference standards to identify skin temperature abnormalities of the upper limbs. However, it is important to consider various confounding factors, and further research is required to validate the accuracy of our results under pathological conditions.(1) Background: although digital infrared thermographic imaging (DITI) is used for diverse medical conditions of the upper limbs, no reference standards have been established. This study aims to establish reference standards by analyzing DITI results of the upper limbs. (2) Methods: we recruited 905 healthy Korean adults and conducted thermography on six regions . We analyzed the data based on the proximity of regions of interest (ROIs), sex, and age. (3) Results: the average temperature (°C) and temperature discrepancy between the right and the left sides (ΔT) of each ROI varied significantly ( Digital infrared thermographic imaging (DITI) has been utilized as an ancillary diagnostic method for various medical conditions related to the upper extremities. For instance, cervical radiculopathy; peripheral nerve entrapment syndrome ; rheumatic disease ; complex regional pain syndrome; tendinopathy; hand arthritis ; and skin cancer can be confirmed by DITI ,11,12,13Consequently, previous attempts to establish reference standards for DITI using systematic reviews based on meta-analysis or machine learning methods have been limited in providing detailed standard DITI values ,16,17,18http://cris.nih.go.kr, accessed on 1 April 2023, number KCT0006880) [This multi-center, single-arm, open label study was conducted in compliance with global/local ethics. The Institutional Review Board of three hospitals approved the study protocol. In addition, this clinical trial was registered with the Clinical Research Information Service of Korea  age between 20 and 69; (2) the absence of any potential contraindication for DITI ; (3) the ability to maintain posture during the test; and (4) the absence of any other issue considered by the investigator to adversely influence DITI results [To minimize potential confounders, the following criteria were adopted: (1) no specific medical history, such as cervical spine disease, diabetes mellitus, peripheral neuropathy or entrapment, joint disorder of the arm or hand, previous history of cervical spine or upper extremity surgery; and (2) currently no pain or skin abnormality of the upper extremity. Initially, 922 healthy Korean adults were enrolled. However, 17 patients were excluded due to test failure or withdrawal of consent. Therefore, the results from 905 participants were included in the analysis . All tesDITI testing was performed using the same equipment and methodology as we previously described . The IriDuring the DITI imaging, upper limbs were divided into six sections based on empirical classification, which facilitated classification into various two-dimensional planes, including the front, back, and lateral sides. The upper limbs were tested in a neutral standing position, encompassing six regions, including the dorsal, ventral, lateral, and medial arm regions, as well as the dorsal and ventral hand regions. Regions of interest (ROIs) were manually marked on both sides and comprised 18 ROIs in the dorsal arm region, 19 ROIs in the ventral arm region, 19 ROIs in the lateral arm region, 7 ROIs in the medial arm region, 26 ROIs in the dorsal hand region, and 26 ROIs in the ventral hand region . Five cet-test, or linear regression analysis were applied according to the purpose of analysis. A p value of less than 0.05 was considered statistically significant.SPSS version 27.0  was used for statistical analysis. Data were checked for normality using the Kolmogorov–Smirnov test, and the results were expressed as mean ± standard deviation (SD), mean with 95% confidence intervals (CI), or median with range. One-way analysis of variance (ANOVA), paired n = 905) was 42.86 ± 12.87 years, and the male-to-female ratio was 411:405. The sample consisted of 183 individuals  in their 20s, 213  in their 30s; 228  in their 40s; 177  in their 50s; and 104 individuals  in their 60s [The average age of all subjects  of each ROI were as follows: 29.22 ± 5.71  and 0.06 ± 0.49  in the dorsal arm region; 29.63 ± 5.74  and 0.01 ± 0.49  in the ventral arm region; 28.45 ± 5.71  and 0.15 ± 0. 62  in the lateral arm region; 28.84 ± 5.57  and 0.04 ± 0.48  in the medial arm region; 29.74 ± 5.14  and 0.06 ± 0.59  in the dorsal hand region; and 28.62 ± 5.44  and 0.07 ± 0.69  in the ventral hand region .p < 0.001, ANOVA)  . p < 0.001, ANOVA). In particular, the |ΔT| in the lateral arm region was significantly larger  than that of any other regions ; ventral arm region temperature = 29.915 − (0.070 × ROIs) ; lateral arm region temperature = 28.684 − (0.063 × ROIs) ; medial arm region temperature = 29.591 − (0.188 × ROIs) ; dorsal hand region temperature = 31.474 − (0.399 × ROIs) ; and ventral hand region temperature = 29.855 − (0.284 × ROIs)   (= 0.429) .p = 0.535, R2 = 0.024); ventral arm region |ΔT| = 0.031 + (0.013 × ROIs) ; lateral arm region |ΔT| = 0.299 − (0.030 × ROIs) ; medial arm region |ΔT| = 0.090 − (0.001 × ROIs) ; dorsal hand region |ΔT| = 0.157 − (0.012× ROIs) ; and ventral hand region |ΔT| = 0.078 + (0.008 × ROIs)   (= 0.061) .p < 0.001, paired t-test). The average temperatures for each region were as follows: 30.31 ± 5.43  for men and 28.32 ± 5.94  for women in the dorsal arm region; 30.72 ± 5.47  for men and 28.73 ± 5.97  for women in the ventral arm region; 29.66 ± 5.45  for men and 27.44 ± 5.92  for women in the lateral arm region; 29.84 ± 5.33  for men and 28.00 ± 5.78  for women in the medial arm region; 30.51 ± 4.93  for men and 28.81 ± 5.38  for women in the dorsal hand region; and 29.71 ± 5.22  for men and 27.72 ± 5.63  for women in the ventral hand region  to 3.80 °C , depending on region and age group .p < 0.001, ANOVA). However, subjects in their 20s had a higher surface temperature than those in their 30s . Consequently, surface temperatures were lowest for those in their 30s for all regions, except for the dorsal hand region   .The average surface temperature of the upper limbs varied depending on regions and ROIs, ranging from 27.08 ± 5.14 °C to 31.27 ± 5.53 °C, which is consistent with the results found in our previous study on the lower limbs. However, the average temperatures of the upper limbs were higher than those of the lower limbs  . We beliThe average of |ΔT| of each ROI also varied by regions and ROIs, ranging from 0.00 to 0.52 °C. However, these values are smaller compared to the reference standards of the lower limbs, where |ΔT| reached 0.76 °C . NeverthThese data for each ROI provide reference standards for DITI of the upper limbs. Clinically significant cold/hot areas, or specific areas with a significant difference between both sides, can be detected based on comparative analysis between the practical patient’s image and these data. Additionally, the detailed ROIs in this study can be modified simply during the processing of DITI capture in actual clinical practice. However, accurate diagnosis requires a comparison of each ROI, not just the averages of whole regions, as normal ranges of surface temperature and ΔT values vary depending on region or ROI.Various intrinsic factors, such as sex, age, fat percentage, and menstrual cycle stage, and extrinsic factors, such as test environment, testing time, and season, can influence the results ,27,28,29A significant correlation was found between the proximity of ROIs to the body core and surface temperature in all regions, except for the lateral arm region, i.e., surface temperatures decreased from the proximal regions to the distal ends. These findings are consistent with a previous suggestion that the surface temperatures of peripheral regions  tend to be cooler than central regions , due to the greater distances from the main thermal organs, such as the heart, large vessels, or viscera ,21,22,23However, the |ΔT| values showed no specific trend based on proximity of ROIs to the body core, which is different from a previous suggestion that the |ΔT| values are higher in the distal regions compared to the proximal regions ,15. ThisSurface temperatures were found to be higher for men in all regions, which is consistent with suggestions that surface temperatures are lower for women due to the insulating effect of thicker subcutaneous fat . This trSurface temperatures increased significantly with age from the 30s, except for the dorsal hand region, which is also consistent with previous research on the lower extremities . These tEstablishing reference standards for DITI is a complex task due to the influence of various confounding factors on skin temperature and DITI measurements . TherefoNonetheless, the study holds value due to its large sample size and the use of a consistent protocol for measurements. It represents the first attempt to establish reference standards for DITI measurements of the upper limbs.The findings of this study provide a basis for establishing reference standards for DITI measurements of surface temperatures in the upper extremities. These standards can aid physicians in making objective diagnoses by comparing patient DITI results with the provided data. However, it is crucial to consider the influence of various confounding factors in surface temperature measurements. Moreover, before confirming the results, it is important to consider several parameters, including the specific location of the ROIs, sex, and age.


Assessment of quality of life (QoL) in patients with tuberculosis (TB) may improve healthcare providers’ understanding of the disease burden. This study aimed to investigate the QoL of patients with TB in Alexandria, Egypt.This cross-sectional study was conducted in chest clinics and main chest hospitals in Alexandria, Egypt. A structured interview questionnaire was used to collect data from participants through face-to-face interviews from November 20, 2021, until the June 30, 2022. We included all adult patients aged 18 years or above during the intensive or continuation phase of treatment. The World Health Organization (WHO) WHOQOL-BREF instrument was used to measure QoL, which includes the physical, psychological, social relationships, and environmental health domains. Using propensity score matching, a group of TB free population was recruited from the same setting and completed the questionnaire.P < 0.0001. Patients with TB aged 18–30 years had the highest environmental score compared with the other age groups (P = 0.021).A total of 180 patients participated in the study: 74.4% were males, 54.4% were married, 60.0% were 18–40 years old, 83.3% lived in urban areas, 31.7% were illiterate, 69.5% reported insufficient income, and 10.0% had multidrug-resistant TB. The TB-free population group had higher QoL scores than the TB patients’ group: (65.0 ± 17.5 vs. 42.4 ± 17.8) for the physical domain, (59.2 ± 13.6 vs. 41.9 ± 15.1) for the psychological domain, (61.8 ± 19.9 vs. 50.3 ± 20.6) for the social domain, (56.3 ± 19.3 vs. 44.5 ± 12.8) for the environment domain, (4.0(3.0–4.0) vs. 3.0(2.0–4.0)) for general health, and (4.0(3.0–4.0) vs. 2.0(2.0–3.0)) for the general QoL, TB had a significant negative impact on QoL, with the physical and psychological domains being the most affected. This finding necessitates strategies to improve QoL of patients with to enhance their compliance to treatment.The online version contains supplementary material available at 10.1186/s12913-023-09381-z. Tuberculosis (TB) is a serious infection caused by a bacterium called Mycobacterium tuberculosis, which typically affects the lungs and can also affect other parts of the body, such as the kidney, spine, or brain. TB is considered one of the top 10 causes of death globally, caused approximately 1.6 million deaths in 2021 [Mycobacterium tuberculosis is thought to infect more than two billion individuals  . AccordiTB is an airborne disease transmitted through the air from the infected person while coughing or speaking. ExposureIndeed, TB control services focus on optimizing microbiological cure and using this parameter as an indicator of treatment success. Although this is critical from a public health perspective, it does not effectively explain the physical, mental, and social suffering of patients with TB . TB can QoL, health related quality of life (HRQoL), and health are sometimes used interchangeably . BowlingWe hypothesized that consideration of TB patients’ QoL during different stages of treatment and the disease process will enable better insight into the patients’ needs, which will help in designing a tailored TB intervention program. Several studies have been performed worldwide on patients with TB to determine their QoL , 11. HowThis cross-sectional study was conducted at chest clinics and hospitals in the Alexandria Governorate. Alexandria is the second-largest city after Cairo and is the largest city on the Mediterranean coast. The city stretches for approximately 40 km (25 miles) along the northern coast of Egypt. It is the fourth largest city in the Arab world, Africa’s ninth largest city, and the world’s 79th -largest urban area by population. In 2022, Alexandria had a population of approximately 5.3 million people. The cluster sample design was considered to increase the precision and representation. The cluster represents the geographic area that contains the TB health facility .TB patients in Egypt are treated at the hospitals and chest clinics of the Ministry of Health and Population (MoHP). Alexandria has nine governmental health facilities where TB-Laboratory tests are carried out and the drug is supplied. They are consisting of seven chest clinics , and two main chest hospitals .Following the calculation of the sample size and determining the total number of clusters, we recorded a list of diagnostic and treatment units (DTU) for each region (administrative zones). Then we designed a random cluster sampling technique, in which both chest clinics and hospitals were sampled randomly in a stepwise manner. Finally, we randomly selected five units to conduct the study, which are: Mamoura chest hospital, Mamoura chest clinic, Bakos chest clinic, Amerya chest clinic, and Kabary chest clinic). The study participants were recruited consecutively from study settings until reaching the required sample size.To calculate the QoL of TB free patients we recruited TB-free population from the same study settings to allow bias-free comparison of QoL scores between TB patients and TB- free population. The size of the matched sample obtained by propensity score matching (PSM) was 131. Using G power software, the calculated power of this study was 99.0% as the mean total QoL mean score of patients with TB patients 45.4 ± 14 (obtained from this current study) and the mean total QoL mean score of the normal population 60.0 ± 14.0 .The sample size of patients with TB was calculated by assuming that the incidence of TB in Egypt is 12 cases per 100,000 people  and the Eligible participants included adult patients with TB, aged 18 years and older, at any TB-treatment phase , and attended the sampled facilities during the study period. We excluded patients with associated pulmonary diseases such as lung cancer, chronic obstructive pulmonary disease, and asthma. Patients with mental disabilities, deaf, and mute were excluded as well.Structured interviewing questionnaire was used to collect data from participants from November 20, 2021, till the end of June 2022 through face-to-face interview. The interview was conducted at the health facility and took 5–10 min. The research team conducted the interview in a separate room / space to ensure patient privacy. The interviewers have already received an educational session on infection control measures. They conducted the interview in a well-ventilated room and wore an N-95 respirator mask. The questionnaire consisted of four sections; the first section was completed from the patient profile to ensure if he/she was eligible or not. Data collectors gathered the following data from patients’ profile: date of symptom onset, date of diagnosis, treatment phase (intensive or continuation phase), current phase start date, (multidrug resistant (MDR) vs. drug sensitive (DS)). The second section was personal and demographic data including . Patients were categorized into low (scored < 40.0%), middle (scored 40.0% to < 70.0%), and high income (scored ≥ 70.0%). The third section included socioeconomic level related questions including .. The est2 test statistics, except for the age groups that turned significant after matching. Though matched groups were considered balanced as this was the best balance available by the sample obtained, and these matched groups were used for purposes of comparisons only between TB patients and TB-free population to allow for the clear highlighting of the difference caused by TB disease and were not used for associations analysis. The balance hypothesis was supported by the graphs Several combinations of confounders which were thought to be of great impact were used for matching and the best results were obtained by using these 4 confounders: age groups, gender, presence of chronic disease other than TB, and residence. Supplementary Table P < 0.05. Statistical analysis was performed using SPSS software version 25 software.Descriptive analysis was used to report the demographic characteristics of the participants. Categorical variables were presented as percentages and continuous variables as means ± standard deviation . We usedP < 0.0001). The scores of the four domain of the TB free population group were significantly higher than that of TB patients’ group. (65.0 ± 17.5 vs 42.4 ± 17.8) for the physical domain, (59.2 ± 13.6 vs 41.9 ± 15.1) for the psychological domain, (61.8 ± 19.9 vs 50.3 ± 20.6) for the social domain, and (56.3 ± 19.3vs 44.5 ± 12.8) for the environment domain respectively. P < 0.0001. Similarly, the median(IQR) scores of general health and the general QoL, were significantly higher among the TB free patients compared with patients with TB (4.0(3.0–4.0) vs 3.0(2.0–4.0), and 4.0(3.0–4.0) vs 2.0(2.0–3.0) respectively) (P < 0.0001) , A total of 180 patients participated in the study, among them, 74.4% were males, 54.4% were married, 60.0% were 18.0–40.0 years with a median value of 37.0 29.0–48.0), 83.3% lived in urban areas, 31.7% were illiterate, 69.5% reported that income was not enough, 58.9% had a crowding index of > 2, and 71.7% were of medium socioeconomic class , 10.0% were treated from MDR-TB, 36.7% were suffering from other comorbidities, 68.3% of them reported food insecurity, 69.4% lost their jobs, 15.6 felt stigmatized and exposed to social ostracism, while 5.6% lost their partners   vs. (42.6 ± 16 vs. 43.4 ± 16.8), however, these differences were not statistically significant. The age group between 18 and 30 years had the highest scores in the physical, psychological and environmental domains, the significant difference was only observed for the environmental domain. Compared to married participants, singles had significantly higher QoL scores in all domains. Patients who lived in urban areas had a higher score in the physical domain than those who lived in rural areas (43.6 ± 18.8 vs. 38.5 ± 18.3), while they had lower scores in the social and environmental domains (49.9 ± 21.2 vs53.6 ± 16.2) and (49.9 ± 21.2 vs. 53.6 ± 16.2) respectively. None of these differences were statistically significant. The scores of psychological domains were comparable in both areas (42.9 ± 16.5 vs. 42.4 ± 14.4). Regarding the university graduates’ patients were of the highest scores in the physical and psychological domains (51.5 ± 20.9 and 45.8 ± 15.4 respectively), while ignorant patients had the lowest scores (38.4 ± 18.1), patients who can only read and write had surprisingly the highest scores in the social domains (57.1 ± 12.7). Scores of the environmental domain were comparable with top score to patients who stopped their education at the junior level. Patients of medium socioeconomic class had the greatest scores in all domains). However, these differences were not significant. Regarding health status of the patients, participants of extrapulmonary TB and who in the continuation phase of treatment had better scores of all domains of QoL than pulmonary TB and patients during the intensive phase, yet this difference was not significant. All QoL domain scores were comparable  in patients with disease durations shorter than and longer than 6 months. Patients of MDR TB had higher scores in physical and psychological domains (44.0 ± 20.4 vs. 42.6 ± 18.7) and (43.1 ± 16.8 vs. 42.6 ± 18.7) respectively, while they had the lowest scores in the social and environmental scores comparing to non-MDR-TB (48.6 ± 20.7 vs. 50.7 ± 20.5) and (47.3 ± 13.5 vs. 44.4 ± 12.8), but this was not considered statistically significant. Patients with chronic diseases other than TB had higher scores in all QoL domains than those who suffered from TB especially in environmental and social domains, yet this difference were not statistically significant  and patients at different treatment phases (intensive or continuation phases) with different diagnoses (pulmonary and extrapulmonary TB) and treatment regimens (drug sensitive and drug resistant) through face-to-face interview. We used probability sampling method (cluster design) to ensure the generalization of the findings. However, the study had many limitations including the inherent limitations of the adopted study design  such recall bias, inability to assess causality, and incapability to study the outcome across different time periods. Second, we did not assess QoL at different time points to describe the change across time concurrently with treatment. Third, we included only adults, future studies are required to include children and youth to investigate the effect of TB on their QoL. Fourth, patients with TB were not matched with TB free population regarding age. Finally, due to the ongoing pandemic and its related public health measures we conducted our study in one government, so larger national studies may be crucial to provide more representation of patients with TB in Egypt.In conclusion, most TB patients have unsatisfactory QoL scores in different domains  compared to TB free population. This may highlight the importance of monitoring QoL as a part of the evaluation of response to long term treatment and medical care for patients with TB to ensure case improvement and better treatment compliance.Below is the link to the electronic supplementary material.Supplementary Material 1Supplementary Material 2