Personalized 3D-printed cranial implants for complex cranioplasty using open-source software.

Background: Cranioplasty is a routine neurosurgery treatment used to correct cranial vault abnormalities. Utilization of 3D printing technology in the field of cranioplasty involving the reconstruction of cranial defects emerged as an advanced possibility of anatomical reshaping. The transformative impact of patient-specific 3D printed implants, focuses on their remarkable accuracy, customization capabilities, and enhanced biocompatibility. Methods: The precise adaptation of implants to patient-specific anatomies, even in complex cases we presented, result in improved aesthetic outcomes and reduced surgical complications. The ability to create highly customized implants addresses the functional aspects of cranial defects and considers the psychological impact on patients. Results: By combining technological innovation with personalized patient care, 3D printed cranioplasty emerges as a transformative avenue in cranial reconstruction, ultimately redefining the standards of success in neurosurgery. Conclusion: 3D printing allows an excellent cranioplasty cosmesis achieved at a reasonable price without sacrificing patient outcomes. Wider implementation of this strategy can lead to significant healthcare cost savings.

The Frequency and Impact of Self-Imposed Elimination Diets on the Nutritional Status and Clinical Course of Disease in Children with Inflammatory Bowel Disease.

BACKGROUND AND AIMS: From the patients' perspective, diet has a relevant role in triggering symptoms of inflammatory bowel disease (IBD). There is a lack of prospective studies regarding the diet of children with IBD. The aim of this study was to assess the frequency and impact of self-imposed elimination diets on the nutritional status and clinical course of disease in the pediatric population. METHODS: This was a prospective case-control study that included newly diagnosed patients with IBD and healthy controls (age/sex-matched peers and siblings) over a one-year period. The participants were examined in three categories: (1) anthropometric data and nutritional status; (2) dietary intake, as obtained by a Food Frequency Questionnaire (FFQ); and (3) dietary beliefs and elimination diets, as obtained by a structured questionnaire. RESULTS: Overall, one-hundred and thirty-seven participants were included (twenty-eight with Crohn's disease, sixteen with ulcerative colitis, three with IBD-unclassified, and seventy healthy controls). Only 15% of patients followed the self-imposed elimination diet upon the diagnosis, which increased to 47.6% by the end of the follow-up. The elimination diet did not influence the nutritional status and quality of the diet. Self-imposed elimination diets were not a risk factor for disease relapse. Most of the patients received nutritional counseling during the follow-up. CONCLUSIONS: The number of patients following self-imposed elimination diets had increased during the disease course but with no influence on nutritional status or relapse risk.

Magnetic Resonance Imaging Can Be Used to Assess Sarcopenia in Children with Newly Diagnosed Crohn's Disease.

BACKGROUND: This study aimed to determine the proportion of patients with sarcopenia diagnosed by MRI and compare these results to bioelectrical impedance analysis (BIA). METHODS: Children with newly diagnosed Crohn's disease (CD) who had MRI enterography (MRE) and BIA and had at least 12 months of follow-up were included. Total psoas muscle area (TPMA) and total paravertebral muscle (TPVM) were measured and compared to subjects' lean mass and the lean mass body index (LMBI) was assessed by BIA. RESULTS: 30 newly diagnosed children with CD were included (mean age 14.2 years, 53% male). Sarcopenia was found in 13 (43%) children; mean TPMA was 15.2 (1.1 SD) cm2 and TPVM 30.95 (1.7 SD) cm2. A highly positive correlation was shown for lean mass assessed by BIA and TPMA (0.706, p < 0.001) and TPVM (0.75, p < 0.001). Sarcopenia was more prevalent in boys (77% vs. 24%, p = 0.004), patients with the perianal disease (69% vs. 29%, p = 0.03), and children with sarcopenia were likely to receive anti-TNF (77% vs. 41%, p = 0.05). During the follow-up period, 16 (53%) children experienced a relapse. TPMA (HR 0.99, p = 0.018) and TPVM (HR 0.99, p = 0.031) values were statistically significant risk factors for relapse. CONCLUSION: A high proportion of patients with CD have sarcopenia at the time of the diagnosis. There is a good correlation between muscle mass assessed by MRI and BIA. Because MRI is performed in a great proportion of newly diagnosed CD patients it can also be used to assess the presence of sarcopenia.

SPECIAL CONSIDERATIONS IN PEDIATRIC TRACHEOSTOMY - A NARRATIVE REVIEW.

Surgical tracheostomy is a life-saving procedure performed for emergent or expectant airway compromise. Morbidity in the pediatric population is higher than in adults due to smaller operating field, immaturity of tissues, anatomic specificities of the child's neck, or the presence of craniofacial dysmorphism. The procedure varies among surgeons regarding the position of the skin incision (vertical or horizontal), resection of the subcutaneous adipose tissue and isthmus of the thyroid gland, use of tracheal flaps, and use of maturation or stay sutures. Both early and late complications can be life-threatening, and include accidental decannulation, stomal plugging, bleeding, and difficult ventilating. Consistent tracheostomal care is crucial in avoiding complications. Primary caregivers must be included and educated about proper stomal care. Decannulation failures are common. Prerequisites for safe decannulation include non-dependence on mechanical ventilation and no recent aspiration events, positive endoscopic airway assessment, and successful daytime capping. The role of polysomnography in decannulation protocols is debated. Although seldom performed, tracheostomy is the procedure of choice in a selected group of pediatric patients. The risks and benefits of the procedure must be weighed for each patient. The education of medical personnel and caregivers is key to reducing serious complications.

From Chilblains (Pernio) to Coeliac Disease-Should We Still Consider It Random?

Coeliac disease (CD) is a gluten-triggered, immune-mediated inflammatory disease occurring in genetically predisposed individuals, causing a variety of gastrointestinal and extraintestinal symptoms. The most common cutaneous association of CD is dermatitis herpetiformis, although recent reports have sought to link CD with other dermatological and autoimmune diseases. Chilblain, also called pernio, is usually a benign, superficial and localized inflammatory skin disorder that results from a maladaptive vascular response to non-freezing cold. We present a patient with pernio (chilblains) and newly diagnosed CD, with a significant intestinal lesion-total villous atrophy, as there are only two known cases of this feature associated with CD published in the literature. In the workup of chilblains (pernio) in children, an active case finding for coeliac disease should be conducted with coeliac-specific serology testing.

Osteomyelitis of the Jaw in COVID-19 Patients: A Rare Condition With a High Risk for Severe Complications.

Osteomyelitis of the jaw is an uncommon infection that arises from the flora of the oral cavity or sinuses and affects immunocompromised and polymorbid patients. Treatment includes surgical debridement and long regiments of broad-spectrum antibiotics. We present three cases of complicated jaw osteomyelitis presented with concurrent COVID-19 infection, including only two reported cases of odontogenic COVID-related osteomyelitis. The two mandibular cases were patients in their 30s with no comorbidities. The first case was an asymptomatic COVID-19-positive patient who developed an odontogenic infection after tooth extraction that was complicated by the second bout of abscess formation and localized osteomyelitis. The second case was a COVID-19-positive patient with an odontogenic infection that presented as airway compromise due to trismus and neck edema, which required an emergency tracheotomy. He developed osteomyelitis of the mandibular ramus that was reconstructed with a titanium plate. The third case was a polymorbid post-COVID-19 patient who developed a protracted infection of the maxillary sinus that resulted in the loss of an eye, destruction of the maxilla, palate, and parts of nasal cavum, and oronasal incontinence. The defect was reconstructed with a microvascular anterolateral thigh flap. We hypothesize that COVID-19-related immune dysfunction and microvascular changes contributed to osteomyelitis in our patients.

Prevalence of Gestational Diabetes Mellitus and Perinatal Outcomes According to the Old Who Criteria and IADPSG Criteria.

BACKGROUND: Gestational diabetes mellitus (GDM) is the most common metabolic disorder in pregnancy. Pregnancies with GDM have worse outcomes compared to pregnancies with normal glucose tolerance. The objectives of the study were to determine the prevalence of GDM and perinatal outcomes according to the old WHO criteria and IADPSG criteria. SUBJECTS AND METHODS: A retrospective study included 2,405 pregnant women who delivered between January 2009 and December 2010. According to the OGTT results, pregnant women were divided into 4 groups. We analyzed the prevalence of GDM, characteristics of pregnant women and their newborns and perinatal outcomes. RESULTS: We found significantly higher prevalence of GDM according to the IADPSG criteria compared to the WHO criteria. Pregnant women with GDM were significantly older, had higher pre-pregnancy BMI, fasting and 2-h plasma glucose. Pregnant women with GDM had worse pregnancy outcomes compared to control group. The overall proportion of overweight and obese pregnant women was the highest in the group of untreated pregnant women with GDM according to the IADPSG criteria. In this group we found significantly higher rate of fetal macrosomia and LGA. The rate of caesarean section was significantly higher in comparison to control group. Pre-eclampsia was significantly more common in groups of pregnant women with GDM compared to control group. CONCLUSION: IADPSG diagnostic criteria reveals more women with hyperglycemic disorders in pregnancy. A group of pregnant women who were normoglycemic according to the WHO criteria, but according to the IADPSG were diagnosed GDM, had adverse pregnancy outcomes. Lower values of glycemia, than those defined for diabetes in pregnancy, are associated with adverse pregnancy outcomes.

Development and Validation of a Food Frequency Questionnaire for Population of Adolescents in Croatia.

RESEARCH BACKGROUND: Food frequency questionnaire (FFQ) is an important method for the estimation of dietary intake in epidemiologic studies. The aim of the study is to develop a FFQ and evaluate its relative validity for adolescents 12 to 18 years old. EXPERIMENTAL APPROACH: FFQ was developed from a previously validated youth/adolescent diet questionnaire (YAQ) by modifying it in order to include Croatian national foods. The final version of the FFQ (FFQ-m) comprised 87 food items. The reference method was a set of two 3-day food records (3DFR) administered twice during the 3 non-consecutive days, one month apart. The FFQ-m was administered approximately on the last day of the second applied dietary food record. Adolescents were recruited from randomly selected elementary and high schools in urban and rural areas of Croatia. FFQ-m was validated on a sample of 84 adolescents (70.2% female). Nutritional intake from FFQ-m and 3DFR were analysed for each participant. Spearman correlation coefficients (r) and Bland-Altman method were used to assess the validity of the FFQ-m compared to 3DFR. Anthropometric parameters were assessed in 78 adolescents. RESULTS AND CONCLUSIONS: The mean nutrient intake estimated by the FFQ-m was higher than that of the 3DFR. The average correlation coefficient for energy and nutrients in our validation study was 0.40. On average, 76.5% of adolescents were classified in the same or adjacent quartile of the nutrient intake. Bland-Altman analysis showed good agreement with all macronutrients and some micronutrients (sodium, phosphorus, potassium, calcium, magnesium and iron). A simple self-administered questionnaire completed by adolescents is a valid tool for measuring energy and nutrient intake among adolescent population. NOVELTY AND SCIENTIFIC CONTRIBUTION: This is the first FFQ developed and validated for population of adolescents in Croatia. It will contribute to further research of nutritional intake in the population of adolescents, especially those from the region.

Workload changes during the COVID-19 pandemic and effects on the flow of cancer patients in the Maxillofacial Surgery Department.

Aim A SARS Coronavirus 2 (COVID-19) pandemic drastically changed the way the health system works. In Croatia, lockdown measures to curb virus spread lasted from March to May 2020, and all non-essential medical procedures and patients' visits have been cancelled. The study aimed to compare patients' flow and interventions in the surgical department before, during and after the lockdown period. Methods This cross-sectional study analysed the workload at the Maxillofacial and Oral Surgery Department (Department), Osijek University Hospital, during the COVID-19 pandemic (March-May 2020) and four subsequent months. The same period of 2019 was compared as a control. The data were subtracted from hospitals' electronic database. Results During COVID-19 lockdown from March to May 2020 the number of hospitalizations (306 vs. 138), surgical procedures (306 vs. 157), and scheduled outpatient visits (2009 vs. 804), dropped significantly as compared to 2019. The number of skin tumour removals was halved (from 155 in 2019 to 58 in 2019) (p<0.001), and the number of emergency patients was unchanged in the 3-month period. A significant decrease in outpatient visits and hospital admissions continued after the lockdown (p<0.001). Conclusion A decrease in the number of outpatient visits, hospitalizations, and tumour removals may result in larger proportions of patients with advanced cancers in the future. The second wave of COVID-19 pandemic is ongoing, and special effort must be paid to reduce the number of cancer patients receiving suboptimal treatment.

Functional Characterization of the γ-Aminobutyric Acid Transporter from Mycobacterium smegmatis MC2 155 Reveals Sodium-Driven GABA Transport.

Characterizing the mycobacterial transporters involved in the uptake and/or catabolism of host-derived nutrients required by mycobacteria may identify novel drug targets against tuberculosis. Here, we identify and characterize a member of the amino acid-polyamine-organocation superfamily, a potential γ-aminobutyric acid (GABA) transport protein, GabP, from Mycobacterium smegmatis The protein was expressed to a level allowing its purification to homogeneity, and size exclusion chromatography coupled with multiangle laser light scattering (SEC-MALLS) analysis of the purified protein showed that it was dimeric. We showed that GabP transported γ-aminobutyric acid both in vitro and when overexpressed in E. coli Additionally, transport was greatly reduced in the presence of ß-alanine, suggesting it could be either a substrate or inhibitor of GabP. Using GabP reconstituted into proteoliposomes, we demonstrated that γ-aminobutyric acid uptake is driven by the sodium gradient and is stimulated by membrane potential. Molecular docking showed that γ-aminobutyric acid binds MsGabP, another Mycobacterium smegmatis putative GabP, and the Mycobacterium tuberculosis homologue in the same manner. This study represents the first expression, purification, and characterization of an active γ-aminobutyric acid transport protein from mycobacteria.IMPORTANCE The spread of multidrug-resistant tuberculosis increases its global health impact in humans. As there is transmission both to and from animals, the spread of the disease also increases its effects in a broad range of animal species. Identifying new mycobacterial transporters will enhance our understanding of mycobacterial physiology and, furthermore, provides new drug targets. Our target protein is the gene product of msmeg_6196, annotated as GABA permease, from Mycobacterium smegmatis strain MC2 155. Our current study demonstrates it is a sodium-dependent GABA transporter that may also transport ß-alanine. As GABA may well be an essential nutrient for mycobacterial metabolism inside the host, this could be an attractive target for the development of new drugs against tuberculosis.

Correction to: Nutritional status and food intake in pediatric patients with inflammatory bowel disease at diagnosis significantly differs from healthy controls.

The authors of the published original version of the above article wanted to correct the below text in the Abstract section.

Membrane Protein Production and Purification from Escherichia coli and Sf9 Insect Cells.

A major obstacle to studying membrane proteins by biophysical techniques is the difficulty in producing sufficient amounts of materials for functional and structural studies. To overexpress the target membrane protein heterologously, especially an eukaryotic protein, a key step is to find the optimal host expression system and perform subsequent expression optimization. In this chapter, we describe protocols for screening membrane protein production using bacterial and insect cells, solubilization screening, large-scale production, and commonly used affinity chromatography purification methods. We discuss general optimization conditions, such as promoters and tags, and describe current techniques that can be used in any laboratory without specialized expensive equipment. Especially for insect cells, GFP fusions are particularly useful for localization and in-gel fluorescence detection of the proteins on SDS-PAGE. We give detailed protocols that can be used to screen the best expression and purification conditions for membrane protein study.

Nutritional status and food intake in pediatric patients with inflammatory bowel disease at diagnosis significantly differs from healthy controls.

Nutritional status and dietary intake in pediatric-onset inflammatory bowel disease are complex and need to be further explored. Therefore, we have assessed anthropometric measures, body composition, and dietary intake of newly diagnosed pediatric patients, and compared them with healthy controls. This was a prospective cross-sectional study including newly diagnosed patients with inflammatory bowel disease (n = 89) and healthy controls (n = 159). Mean energy intake was significantly lower in healthy controls compared to patients with ulcerative colitis, but not in patients with Crohn's disease. Intake of all macronutrients, dietary fiber, and calcium was significantly lower in patients with ulcerative colitis, whereas the only intake of animal protein, fruit, and calcium differed significantly in patients with Crohn's disease. There were no significant differences in the body fat percentage between patients with ulcerative colitis or Crohn's disease vs. controls; however, lean mass-for-age z-scores were significantly lower in patients with both diseases in comparison to controls.Conclusion: Food intake of newly diagnosed pediatric patients with inflammatory bowel disease significantly differed from healthy controls. Altered anthropometry and body composition are present already at the time of diagnosis. What is Known: • Children with inflammatory bowel disease suffer from malnutrition, especially children with Crohn's disease in whom linear growth failure often precedes gastrointestinal symptoms. What is New: • This study showed significantly lower intake of energy, macronutrients, and various micronutrients in patients with ulcerative colitis compared to healthy controls, while patients with Crohn's disease have a lower intake of fruits, calcium, and animal protein at diagnosis. • Altered body composition is present in both groups of patients at the time of diagnosis.

Glutamate transporters: a broad review of the most recent archaeal and human structures.

Glutamate transporters play important roles in bacteria, archaea and eukaryotes. Their function in the mammalian central nervous system is essential for preventing excitotoxicity, and their dysregulation is implicated in many diseases, such as epilepsy and Alzheimer's. Elucidating their transport mechanism would further the understanding of these transporters and promote drug design as they provide compelling targets for understanding the pathophysiology of diseases and may have a direct role in the treatment of conditions involving glutamate excitotoxicity. This review outlines the insights into the transport cycle, uncoupled chloride conductance and modulation, as well as identifying areas that require further investigation.

Binding of EphrinA5 to RET receptor tyrosine kinase: An in vitro study.

Eph/Ephrin signaling pathways are crucial in regulating a large variety of physiological processes during development, such as cell morphology, proliferation, migration and axonal guidance. EphrinA (efn-A) ligands, in particular, can be activated by EphA receptors at cell-cell interfaces and have been proposed to cause reverse signaling via RET receptor tyrosine kinase. Such association has been reported to mediate spinal motor axon navigation, but conservation of the interactive signaling pathway and the molecular mechanism of the interaction are unclear. Here, we found Danio rerio efn-A5b bound to Mus musculus EphA4 with high affinity, revealing structurally and functionally conserved EphA/efn-A signaling. Interestingly, we observed no interaction between efn-A5b and RET from zebrafish, unlike earlier cell-based assays. Their lack of association indicates how complex efn-A signaling is and suggests that there may be other molecules involved in efn-A5-induced RET signaling.

Comparison of Obesity Prevalence and Dietary Intake in School-Aged Children Living in Rural and Urban Area of Croatia.

The aim of this study was to explore the differences in nutritional status and dietary intakes in 12~17 year-old children living in urban (Zagreb) and the rural (Sinj and Drnis) area of Croatia. A validated food frequency questionnaire was used to assess dietary intake. For each participant, body weight and body height were measured and bioelectrical impedance was used to estimate the body fat percentage. There was an overall of 310 children included: 105 (33.9%) from urban area and 205 (66.1%) from rural area; 191 (61.6%) were female with a mean age 14.9 (range 12~17) years. When adjusted for age and gender, there was no statistically significant difference in body mass index for age Z-scores between urban and rural parts (0.23±0.07 vs. 0.30±1.15; P=0.650) or in average daily energy intake (2,479.2±1,111.2 kcal vs. 2,338.2±920.2 kcal; P=0.702). There was a statistically significant difference in nutritional status between genders, with a higher percentage of boys being overweight or obese compared to girls. When combined, 'Fast food' and 'Snacks' were major contributors to the total energy intake for both areas. The mean contribution of 'Fast food' to total energy intake was significantly higher in the urban area. The prevalence of obesity among Croatian children is high and unrelated to the urban/rural setting, which could be partially explained by the high intake of 'Fast food' and 'Snacks'.

Bifidobacterium animalis subsp. lactis in prevention of common infections in healthy children attending day care centers - Randomized, double blind, placebo-controlled study.

BACKGROUND & AIMS: The aim of our study was to investigate the role of Bifidobacterium animalis subsp. lactis (BB-12(®)) in the prevention of common (gastrointestinal and respiratory) infections in healthy children who attend day care centers. METHODS: We conducted a randomized, double-blind, placebo-controlled trial in 210 children who attend day care centers. They were randomly allocated to receive placebo (Placebo group, n = 106) or BB-12(®) at a dose of 10(9) colony-forming units (CFU) (Intervention group, n = 104) during the 3-month intervention period. RESULTS: Intention to treat analysis was used. There were overall 99 infections in Placebo group and 97 in Intervention group (incidence rate ratio = 1.0014, p = 0.992, Poisson regression model). Overall 65 children (61.3%) in Placebo group and 67 (64.4%) in Intervention group had common infections (p = 0.642). Mean number of infections per child was 0.93 (range 0-3) in Placebo group and 0.93 (range 0-3) in Intervention group (p = 0.898). There was no difference in secondary (duration of symptoms, number of children with gastrointestinal and respiratory tract infections, absence from day care center due to infections, use of antibiotics) and exploratory (type of gastrointestinal and respiratory tract infection) endpoints between groups. CONCLUSION: Results of performed study show that BB-12(®) has no effect on the prevention of gastrointestinal and respiratory tract infections in healthy children who attend day care centers.

Methotrexate is an efficient therapeutic alternative in children with thiopurine-resistant Crohn's disease.

OBJECTIVE: This study aimed to investigate the role of methotrexate (MTX) in the maintenance of clinical remission and mucosal healing in children with Crohn's disease (CD), in whom azathioprine (AZA) treatment failed. MATERIALS AND METHODS: This was a retrospective, longitudinal cohort study which included all children who were diagnosed with CD during a period of 10 years and who received MTX for ≥12 months after failed AZA treatment. Remission was assessed clinically, defined by Pediatric Crohn's Disease Activity Index as a score of ≤10 and no need for the reintroduction of the remission induction therapy. In the subset of patients with sustained clinical remission, the rate of mucosal healing was endoscopically assessed. Endoscopic lesions were assessed by Simple Endoscopic Score for CD. Each patient served as his or her own historical control. RESULTS: Of the 32 included patients, 22 (68.7%) remained in the stable clinical remission after a period of 12 months and 14 (43.8%) did not experience relapse during the whole follow up (median duration 2.9 years; range 1-4.8 years). From all patients who were in clinical remission during the entire follow up (n = 14), endoscopy was performed in eight (57%) patients and showed complete mucosal healing macroscopically (Simple Endoscopic Score for CD score of 0) and microscopically in seven out of eight (87.5%) patients. CONCLUSION: MTX was found to be an efficient therapeutic alternative in the thiopurine-resistant patients, enabling the complete mucosal healing.