Advanced and Safe Synthetic Microbial Chassis with Orthogonal Translation System Integration.

Through the use of CRISPR-assisted transposition, we have engineered a safe Escherichia coli chassis that integrates an orthogonal translation system (OTS) directly into the chromosome. This approach circumvents the limitations and genetic instability associated with conventional plasmid vectors. Precision in genome modification is crucial for the top-down creation of synthetic cells, especially in the orthogonalization of vital cellular processes, such as metabolism and protein translation. Here, we targeted multiple loci in the E. coli chromosome to integrate the OTS simultaneously, creating a synthetic auxotrophic chassis with an altered genetic code to establish a reliable, robust, and safe synthetic protein producer. Our OTS-integrated chassis enabled the site-specific incorporation of m-oNB-Dopa through in-frame amber stop codon readthrough. This allowed for the expression of advanced underwater bioglues containing Dopa-Lysine motifs, which are crucial for wound healing and tissue regeneration. Additionally, we have enhanced the expression process by incorporating scaffold-stabilizing fluoroprolines into bioglues, utilizing our chassis, which has been modified through metabolic engineering (i.e., by introducing proline auxotrophy). We also engineered a synthetic auxotroph reliant on caged Dopa, creating a genetic barrier (genetic firewall) between the synthetic cells and their surroundings, thereby boosting their stability and safety.

Features that characterize monoclonal light chain ("myeloma") cast nephropathy with immunofluorescence challenges and emphasis on electron microscopy.

Renal disease is a common cause of morbidity and mortality in patients with plasma cell dyscrasias. The serum-free light chain assay is used in patients, mostly older, with unexplained acute kidney injury to screen for potential myeloma cast nephropathy. This study consists of a systematic review of diagnostic features in myeloma cast nephropathy. The morphological features of tubular casts in patients with multiple myeloma have not been systematically analyzed. This study focuses on the morphology of these casts, emphasizing ultrastructural features, in a series of 23 patients with light chain ("myeloma") cast nephropathy and compared them with casts in 10 patients with various diseases. The immunofluorescence data were correlated with morphological findings to provide diagnostic assessments and practice guidelines. The ultrastructural features identified as diagnostic of casts associated with myeloma included: amyloid and crystals in the casts, multiple well-defined fracture planes forming a complex jigsaw puzzle arrangement of cast contents, indicative of the fragility of the immunoglobulin light chains involved, and reactive tubular cells lining the tubules with the casts. These features were seen in 95.2% of MCN cases and none of the casts in other renal conditions. Myeloma casts exhibited light chain monoclonality in a significant percentage of the MCN cases and often no staining for IgA or IgM. In contrast, the majority of non-myeloma casts stained for both kappa and lambda light chains, lgA, and lgM, and showed ultrastructurally a rather uniform finely to coarsely granular electron density occasionally admixed with cellular debris.

Refractory type 1 plastic bronchitis in a child; case report.

BACKGROUND: Plastic bronchitis (PB) is a rare pediatric pulmonary condition characterized by the production of branching bronchial casts that cause partial or total obstruction of the bronchial lumen. CASE PRESENTATION: We describe a 13-year-old boy with a history of bronchial asthma and left lower lobectomy, with persistent cough and left-sided chest pain when he went to the emergency room. Chest radiography showed complete left lung opacity denoting total left lung collapse, and flexible bronchoscopy revealed cohesive casts totally occluding the left bronchus, with frequent recurrence that finally ended with left pneumonectomy. CONCLUSION: Plastic bronchitis is a rare, fatal disease in children that requires a high index of suspicion for both diagnosis and treatment. Although bronchoscopic removal of the bronchial casts together with the medical treatment are the main lines of treatment, cases with recurrent formation of casts are at high risk for surgical intervention in the form of either lobectomy or pneumonectomy.

Feasibility of composted green waste amended by vermiculite and earthworm casts as the growth media for three common ornamental plants.

This study demonstrated the effects of adding specific proportions of vermiculite (VMT: 0%, 10%, and 20%) and earthworm casts (EWCs: 0%, 10%, and 20%) on the physico-chemical properties of composted green waste (CGW), and the impacts of amended CGW as growth media on the growth of three common ornamental plants (Dahlia pinnata Cav. [dahlia], Centaurea cyanus L. [cornflower], and Consolida ajacis [L.] Schur [delphinium]). Compared with Treatment T1 (CK), the addition of 10% VMT and 20% EWCs greatly (p < 0.05) increased the total porosity, aeration porosity, water-holding porosity, total nitrogen, available phosphorus, available potassium, and organic matter of CGW by 9%, 35%, 4%, 18%, 27%, 13%, and 33%, respectively. In addition, this pattern increased (p < 0.05) the total fresh biomass, total chlorophyll content, and root length of dahlias by 9%, 19%, and 27%, respectively; those of cornflowers by 17%, 30%, and 29%, respectively (p < 0.05); and those of delphiniums by 23%, 14%, and 63%, respectively. Therefore, the amended CGW supplemented with 10% VMT and 20% EWCs was an ideal growth medium for the three plants.

Morphological and morphometric study of the hominin dental casts from Grotta-Riparo di Uluzzo C (Apulia, southern Italy).

OBJECTIVES: Grotta-Riparo di Uluzzo C (Apulia, southern Italy) is a pivotal site for investigating the evolution of the Middle Paleolithic and the earliest phases of the Upper Paleolithic in southern Italy, as the extensive stratigraphic record of this site includes a thick Mousterian sequence followed by the Uluzzian. Here, we investigate the taxonomic affinity of seven unpublished deciduous human teeth retrieved from the site of Uluzzo C in 1960. MATERIALS AND METHODS: The teeth are represented by seven plaster dental casts, which are housed at the Museo Civico di Paleontologia e Paletnologia in Maglie (Lecce, Apulia). The location of the original specimens remains unknown, rendering these casts the only human remains evidence yielded by Uluzzo C to date. Based on occlusal-view photographs and digital models of the casts, we examined the external morphology and morphometry of the teeth, comparing them to Homo sapiens and H. neanderthalensis samples. Through geometric morphometric methods and statistical analyses, we analyzed the crown outline of the deciduous molars. RESULTS: The teeth show morphological and morphometric features that are variably found in H. neanderthalensis, H. sapiens, or both. Specifically, crown outline analysis shows that all molars fall within H. neanderthalensis variability, except for Uluzzo 853 (lower right deciduous first molar), which falls within H. sapiens variability. DISCUSSION: This study provides the first taxonomic assessment of the hominin teeth from Uluzzo C. The results contribute additional insights into the Paleolithic peopling of southern Italy during a crucial period marked by the persistence of post-Tyrrhenian Neanderthal techno-complexes and the arrival of H. sapiens.

Predictability of expansion movements performed by clear aligners in mixed dentition in both arches: a retrospective study on digital casts.

BACKGROUND: to evaluate the predictability of expansion achieved in patients in early mixed dentition treated with Clear Aligners (CA), analyzing the efficiency of the expansion at the end of the first set of aligners and at the end of the therapy in the upper and lower arch. METHODS: 36 patients (20 F, 16 M; mean age 8.3 ± 1.5 years) were selected retrospectively from the Department of Orthodontics of the Hospital of Rome "Tor Vergata". All subjects were treated with CA with no other auxiliaries than attachments. For each patient a standardized sequential expansion protocol was planned for both arches. Digital dental casts were created at three observation periods from an intraoral scanner: prior to treatment (T0), at the end of the first set of aligners (T1), at the end of treatment (T2). The 3D models in planned position determined by the first Clincheck (CC) were obtained for comparison with T1 and T2. Six linear transversal measurements were used to evaluate the dimensional changes and the predictability of expansion movements, comparing T1-CC and T2-CC. RESULTS: a statistically significant increase within the pre-treatment and the final outcomes for all the variables examined was found. In the upper arch, the greatest level of predictability was detected at the level of the first (46.44%) and second deciduous molar width (44.95%) at T1. The analysis of T2-CC changes showed a significant increase in the percentage of predictability of expansion at the level of the first permanent molars, at mesial (54.86%) and distal (58.92%) width. In the lower arch, a higher percentage of predictability than the upper arch was reported at T1-CC and T2-CC, with the greatest values at the level of second (T1-CC: 48.70%; T2-CC: 75.32%) and first deciduous molar width (T1-CC: 45.71%; T2-CC: 72.75%). CONCLUSIONS: CA can induce significant transversal increments. The predictability of expansion is variable, but it did not exceed the 50% during the first set of aligners. It was necessary to apply refinement set to achieve a good predictability for expansion of about 70%. The expansion in the lower arch was observed to be more predictable than in the upper arch.

Comparison of dental dimensions in models developed with digital procedures and plaster models / Comparación de dimensiones dentales en modelos desarrollados con procedimientos digitales y modelos en yeso

Aim: This study aimed to collect evidence on the validity and reliability of measurements obtained from digital impression techniques. Materials and Methods: This comparative study was conducted on 31 patients. Intraoral scanner was applied to all patients. For each patient, an alginate impression of the upper maxilla was taken and later the 3D digital model was extracted by dental cone-beam computed tomography (CBCT). For preparation of plaster models, alginate impressions were taken and immediately poured with dental stone. In the next stage, a comparison was performed among the intraoral scanner, CBCT, and plaster models in terms of tooth size, dental width, and intra-arch dimensions. Results: Measuring tooth size and intra-arch dimensions in digital images obtained from intraoral scanner and CBCT were in most cases lower than the results obtained in the plaster models but the differences between digital techniques and plaster models are not clinically noticeable. Conclusions: Digital systems including intraoral scanner and CBCT are acceptable for clinical use in terms of accuracy.

Objetivo: Este estudio tuvo como objetivo recopilar evidencia sobre la validez y confiabilidad de las mediciones obtenidas a partir de técnicas de impresión digital. Materiales y Métodos: Este estudio comparativo se realizó en 31 pacientes. A todos los pacientes se les aplicó escáner intraoral. Para cada paciente, se tomó una impresión de alginato del maxilar superior y posteriormente se extrajo el modelo digital 3D mediante Tomografía computarizada de haz cónico (CBCT) dental. Para la preparación de los modelos de yeso se tomaron impresiones de alginato y se vertieron inmediatamente con yeso dental. En la siguiente etapa, se realizó una comparación entre el escáner intraoral, CBCT y los modelos de yeso en términos de tamaño de diente, ancho dental y dimensiones intraarcada. Discusión: Se encontró que la apariencia microscópica de las células fusiformes era comparable en ambos grupos. Los resultados de la citometría de flujo demostraron expresiones comparables en ambos grupos, siendo las muestras positivas para CD90, CD73, CD105, HLA ABC y negativas para CD34, CD45 y HLA DR. Hubo variaciones en la expresión de los marcadores cuando se evaluaron los potenciales de diferenciación. Conclusión: Los sistemas digitales como el escáner intraoral y el CBCT son aceptables para uso clínico en términos de precisión.

Flexible and Rigid Bronchoscopy for Critically Ill Children on Extracorporeal Membrane Oxygenation.

BACKGROUND: We aim to describe our experience with bronchoscopy to diagnose and relieve tracheobronchial obstruction in anticipation of decannulation in children on extracorporeal membrane oxygenation (ECMO) support. METHODS: A retrospective cohort study of children on ECMO between 1/2018 and 12/2022. RESULTS: A total of 107 children required ECMO support during the study period for cardiac (n = 48, 45%), pulmonary (n = 38, 36%), or cardiopulmonary dysfunction (n = 21, 20%). Thirty-seven (35%) patients underwent 99 bronchoscopies while on ECMO. Most (76%, n = 75) experienced no improvement or worsening of chest radiography 24 hours following bronchoscopy. Clinical improvement in tidal volumes 48 hours after the first bronchoscopy was noted in 13/25 patients with available data (p = 0.05). Adverse events were seen in 18 (49%) patients who underwent bronchoscopy, including pneumothorax (n = 8, 22%), pneumonia (n = 7, 19%), pulmonary hemorrhage (n = 6, 16%), and sepsis (n = 5, 14%). ECMO courses were longer (25.4 ± 37.2 vs 6.1 ± 8.8 days, p < 0.0001) and more likely to be complicated by pneumonia (p = 0.0004) and sepsis (p = 0.047) in patients who underwent bronchoscopy compared with those who did not. Adverse events following bronchoscopy were associated with the number of bronchoscopies (p = 0.0003) and the presence of obstructive materials but not with the type of bronchoscopy or indication for ECMO. Mortality rates were similar between patients who underwent bronchoscopy and those who did not. CONCLUSION: Children requiring bronchoscopy represent a subset of the sickest children on ECMO. Bronchoscopy may provide benefit in children with persistent cardiopulmonary failure who could not otherwise be decannulated. Adverse events are associated with the number of bronchoscopies and the presence of obstructive material. LEVEL OF EVIDENCE: 4 Laryngoscope, 134:4134-4140, 2024.

Is reduction necessary in overriding metaphyseal distal radius fractures in children under 11 years: a systematic review and meta-analysis of comparative studies.

OBJECTIVE: To determine the necessity of reduction in the treatment of overriding metaphyseal distal radius fractures (DRF) in children under 11 years. METHODS: In this systematic review and meta-analysis, PubMed, Embase, and Cochrane databases were searched to retrieve studies published from inception to 2023. Two reviewers independently screened for studies with observational or randomized control design comparing two treatments for overriding metaphyseal DRF in patients under 11 years: simple casting without reduction (SC group) versus closed reduction plus casting or pin fixation (CRC/F group); with varying outcomes reported (CRD471761). The risk of bias was assessed using the ROBINS-I tool. RESULTS: Out of 3,024 screened studies, three met the inclusion criteria, 180 children (mean age 7.1 ± 0.9 years) with overriding metaphyseal DRF: SC-group (n = 79) versus CRC/F-group (n = 101). Both treatment groups achieved 100% fracture consolidation without requiring further manipulation. The SC-group showed significantly fewer complications (mean difference [MD] 0.08; 95% CI [0.01, 0.53]; I2 = 22%; P < 0.009) and trends towards better sagittal alignment (MD 5.11; 95% CI [11.92, 1.71]; I2 = 94%; P < 0.14), less reinterventions (MD 0.31; 95% CI [0.01, 8.31]; P < 0.48), and fewer patients with motion limitation at the end of follow-up (MD 0.23; 95% CI [0.03,  1.98]; P < 0.18), although these findings were not statistically significant. CONCLUSIONS: Despite a limited number of studies comparing SC versus CRC/F in overriding DRF in children under 11 years, this study suggests that anatomical reduction is not necessary. Treating these fractures with SC, even when presenting with an overriding position, leads to reduced complications, shows a trend towards fewer reinterventions, improved sagittal alignment, and less limitation in patient motion. LEVEL OF EVIDENCE: Level III, Systematic review of Level-III studies.

RNA-guided genome engineering: paradigm shift towards transposons.

CRISPR-Cas systems revolutionized the genome engineering field but need to induce double-strand breaks (DSBs) and may be difficult to deliver due to their large protein size. Tn7-like transposons such as CRISPR-associated transposons (CASTs) can be repurposed for RNA-guided DSB-free integration, and obligate mobile element guided activity (OMEGA) proteins of the IS200/IS605 transposon family have been developed as hypercompact RNA-guided genome editing tools. CASTs and OMEGA are exciting, innovative genome engineering tools that can improve the precision and efficiency of editing. This review explores the recent developments and uses of CASTs and OMEGA in genome editing across prokaryotic and eukaryotic cells. The pros and cons of these transposon-based systems are deliberated in comparison to other CRISPR systems.

3D Digital Modeling of Dental Casts from Their 3D CT Images with Scatter and Beam-Hardening Correction.

Dental 3D modeling plays a pivotal role in digital dentistry, offering precise tools for treatment planning, implant placement, and prosthesis customization. Traditional methods rely on physical plaster casts, which pose challenges in storage, accessibility, and accuracy, fueling interest in digitization using 3D computed tomography (CT) imaging. We introduce a method that can reduce both artifacts simultaneously. To validate the proposed method, we carried out CT scan experiments using plaster dental casts created from dental impressions. After the artifact correction, the CT image quality was greatly improved in terms of image uniformity, contrast-to-noise ratio (CNR), and edge sharpness. We examined the correction effects on the accuracy of the 3D models generated from the CT images. As referenced to the 3D models derived from the optical scan data, the root mean square (RMS) errors were reduced by 8.8~71.7% for three dental casts of different sizes and shapes. Our method offers a solution to challenges posed by artifacts in CT scanning of plaster dental casts, leading to enhanced 3D model accuracy. This advancement holds promise for dental professionals seeking precise digital modeling for diverse applications in dentistry.

Smile aesthetics in Pakistani population: dentist preferences and perceptions of anterior teeth proportion and harmony.

BACKGROUND: This study aimed to evaluate dentist perceptions of attractive smiles in the Pakistani population, considering different dental proportions. METHODS: Maxillary casts and digital images were used to create symmetrical representations of anterior teeth. dentists' preferences for good and bad teeth proportions, width/height ratios, and various dental proportions (golden, recurring esthetic dental (RED), golden percentage, Preston, and local/observed) were assessed using one sample and paired t-test. The Chi-square test was used to determine the gender disparities and factors affecting smile attractiveness. A p-value of ≤ 0.05 was taken as significant. RESULTS: The RED proportion emerged as the preferred choice for normal-sized teeth, with specialists and general dentists favoring it over the golden proportion. For tall teeth, the golden proportion was predominantly preferred. The golden percentage received limited preference for aesthetic smile construction. CONCLUSIONS: The smiles created using the principles of RED proportion were opted as the most attractive by local dentists. Factors such as tooth arrangement, color, and midline were highlighted as essential considerations in aesthetic smile construction.

Mobile genetic element-based gene editing and genome engineering: Recent advances and applications.

Genome engineering has revolutionized several scientific fields, ranging from biochemistry and fundamental research to therapeutic uses and crop development. Diverse engineering toolkits have been developed and used to effectively modify the genome sequences of organisms. However, there is a lack of extensive reviews on genome engineering technologies based on mobile genetic elements (MGEs), which induce genetic diversity within host cells by changing their locations in the genome. This review provides a comprehensive update on the versatility of MGEs as powerful genome engineering tools that offers efficient solutions to challenges associated with genome engineering. MGEs, including DNA transposons, retrotransposons, retrons, and CRISPR-associated transposons, offer various advantages, such as a broad host range, genome-wide mutagenesis, efficient large-size DNA integration, multiplexing capabilities, and in situ single-stranded DNA generation. We focused on the components, mechanisms, and features of each MGE-based tool to highlight their cellular applications. Finally, we discussed the current challenges of MGE-based genome engineering and provided insights into the evolving landscape of this transformative technology. In conclusion, the combination of genome engineering with MGE demonstrates remarkable potential for addressing various challenges and advancing the field of genetic manipulation, and promises to revolutionize our ability to engineer and understand the genomes of diverse organisms.

A morphometric approach to morphology analysis of palatal rugae in sibling groups.

Recent advances in imaging technologies, such as intra-oral surface scanning, have rapidly generated large datasets of high-resolution three-dimensional (3D) sample reconstructions. These datasets contain a wealth of phenotypic information that can provide an understanding of morphological variation and evolution. The geometric morphometric method (GMM) with landmarks and the development of sliding and surface semilandmark techniques has greatly enhanced the quantification of shape. This study aimed to determine whether there are significant differences in 3D palatal rugae shape between siblings. Digital casts representing 25 pairs of full siblings from each group, male-male (MM), female-female (FF), and female-male (FM), were digitized and transferred to a GM system. The palatal rugae were determined, quantified, and visualized using GMM computational tools with MorphoJ software (University of Manchester). Principal component analysis (PCA) and canonical variates analysis (CVA) were employed to analyze palatal rugae shape variability and distinguish between sibling groups based on shape. Additionally, regression analysis examined the potential impact of shape on palatal rugae. The study revealed that the palatal rugae shape covered the first nine of the PCA by 71.3%. In addition, the size of the palatal rugae has a negligible impact on its shape. Whilst palatal rugae are known for their individuality, it is noteworthy that three palatal rugae (right first, right second, and left third) can differentiate sibling groups, which may be attributed to genetics. Therefore, it is suggested that palatal rugae morphology can serve as forensic identification for siblings.

Anatomical variations in the cerebral arterial circle of the Saimaa (Pusa hispida saimensis) and Baltic ringed seals (Pusa hispida botnica).

The intracranial arterial vascularization of the Saimaa ringed seals (Pusa hispida saimensis; Nordquist, 1899) and Baltic ringed seals (Pusa hispida botnica; Gmelin, 1788) disclosed patterns of anatomical architecture comparable to that of other pinniped species. Arterial silicone casts on skull scaffolds, and magnetic resonance imaging (MRI) showed that the besides joining the caudal communicating arteries upon entering the cerebral arterial circle, the bilateral internal carotid arteries bifurcated as laterally oriented rostral choroidal arteries and rostral cerebral arteries. The latter arteries almost immediately gave off the laterally oriented middle cerebral arteries. Numerous individual variations were evident in differences in the exact branching sites of bilateral vessels or the size or number of arterial branches. Two Saimaa ringed seals had only a tiny foramen for the left internal carotid artery to enter the intracranial space, and the intracranial part of this vessel was short. It did not reach the cerebral arterial circle. The intracranial part of the right internal carotid artery is bifurcated and also supplied the left side of the cerebral arterial circle. Both specimens had aplasia of the left rostral cerebral artery. The intracranial arterial arrangement of Saimaa and Baltic ringed seals reflects the arterial architecture of this body region in terrestrial mammals with little evidence for aquatic adaptations or changes related to thermoregulation.

The past, present and future of the conservative treatment of distal radius fractures.

The distal radius fracture is a common fracture with a prevalence of 17% on the emergency departments. The conservative treatment of distal radius fractures usually consists of three to six weeks of plaster immobilization. Several studies show that one week of plaster immobilization is safe for non- or minimally displaced distal radius fractures that do not need reduction. A shorter period of immobilization may lead to a better functional outcome, faster reintegration and participation in daily activities. Due to upcoming innovations such as three-dimensional printed splints for distal radius fractures, a patient specific splint can be produced which may offer more comfort. Furthermore, these three-dimensional printed splints are expected to be more environmental friendly in comparison with traditional plaster casts.

Total contact casts versus removable offloading interventions for the treatment of diabetic foot ulcers: a systematic review and meta-analysis.

Objective: This study aimed to evaluate the effectiveness of total contact casts (TCCs) versus removable offloading interventions among patients with diabetic foot ulcers (DFUs). Methods: A comprehensive search was done in databases Embase, Cochrane Library, and, PubMed. The references of retrieved articles were reviewed, up until February 2023. Controlled trials comparing the effects of TCCs with removable offloading interventions (removable walking casts and footwear) in patients with DFUs were eligible for review. Results: Twelve studies were included in the meta-analysis, involving 591 patients with DFUs. Among them, 269 patients were in the intervention group (TCC), and 322 in the control group (removable walking casts/footwear). The analysis revealed that the TCC group had higher healing rates (Risk Ratio(RR)=1.22; 95% confidence interval(CI):1.11 to 1.34, p<0.001), shorter healing time (Standard Mean Difference(SMD)=-0.57; 95%CI: -1.01 to -0.13, P=0.010), and elevated occurrence of device-related complications (RR=1.70; 95%CI:1.01 to 2.88, P=0.047), compared with the control group. Subgroup analysis illustrated patients using TCCs had higher healing rates than those using removable walking casts (RR=1.20; 95%CI:1.08 to 1.34, p=0.001) and footwear (RR=1.25; 95%CI:1.04 to 1.51, p=0.019), but they required comparable time for ulcer healing compared with those using removable walking casts (SMD=-0.60; 95%CI: -1.22 to 0.02, P=0.058) or footwear group (SMD=-0.52; 95%CI: -1.17 to 0.12, P=0.110). Although patients using TCCs had significantly higher incidence of device-related complications than those using footwear (RR=4.81; 95%CI:1.30 to 17.74, p=0.018), they had similar one compared with those using the removable walking casts (RR=1.27; 95%CI:0.70 to 2.29, p=0.438). Conclusion: The use of TCCs in patients with DFUs resulted in improved rates of ulcer healing and shorter healing time compared to removable walking casts and footwear. However, it is important to note that TCCs were found to be associated with increased prevalence of complications.

A case report: Acute kidney injury with progression to chronicity in an eldery woman.

Acute renal failure in elderly patients can be caused by a wide spectrum of diseases that usually have a cause outside the kidney. The most common causes include renal impairment as part of ANCA vasculitis, another category includes clonal plasmatic cell disease with light chain cast nephropathy; and there also exists an increasing number of drug-induced tubulointerstial damage. We present a case of iatrogenic less common form of acute failure in a 73-year-old woman, who did not suffer from any serious disease until then. Although the biopsy helped to determine the cause of the failure and thus affect subsequent therapy, the function did not return to the previous state and the patient progressed to CKD G3bA1 with serum creatinine values of around 170-140 µmol/l.