NCB-02(standardized Curcumin preparation)protects dinitrochlorobenzene-induced colitis through down-regulation of NFκ-B and iNOS

AIM:To evaluate the efficacy and mechanism of action of NCB-02,a standardized Curcumin preparation,against 2,4-dinitrochlorobenzene(DNCB)-induced ulcerative colitis in rats.METHODS:Ulcerative colitis was induced in male rats by sensitizing with topical application of DNCB in acetone for 14 d and intra-colonol challenge with DNCB on day 15.A separate group of animals with vehicle treatment in similar fashion served as control group.Colitis rats were divided into different groups and treated with NCB-02 at doses of 25,50 and 100 mg/kg b.wt p.o.for 10 d.Sulfasalazine at a dose of 100 mg/kg b.wt for 10 d served as a reference group.On day 10 after respective assigned treatment,all the animals were euthanized and the length of the colon,weight of entire colon and distal 8 cm of the colon were recorded.The distal part of the colon was immediately observed under a stereomicroscope and the degree of damage was scored.Further distal 8 cm of the colon was subject to the determination of colonic myeloperoxidase(MPO),lipid peroxidation(LPO)and alkaline phosphatase (ALP)activities.A small piece of the sample from distal colon of each animal was fixed in 10% neutral buffered formalin and embedded in paraffin wax and sectioned for immunohistochemical examination of NFκ-B and iNOS expression.RESULTS:NCB-02 showed a dose dependent protection against DNCB-induced alteration in colon length and weight.NCB-02 treatment also showed a dose dependent protection against the elevated levels of MPO,LPO and ALP,induced by DNCB.NCB-02 demonstrated a significant effect at a dose of 100 mg/kg b.wt.,which was almost equipotent to 100 mg/kg b.wt.of sulfasalazine.Treatment with sulfasalazine and curcumin at a dose of 100 mg/kg b.wt.inhibited the DNCB-induced overexpression of NFκ-B and iNOS in the colon.CONCLUSION:Curcumin treatment ameliorates colonic damage in DNCB-induced colitic rats,an effect associated with an improvement in intestinal oxidative stress and downregulation of colonic NFκ-B and iNOS expression.     

Hypoxic Regulation of Mitochondrial Metabolism and Mitophagy in Nucleus Pulposus Cells Is Dependent on HIF-1α–BNIP3 Axis

Nucleus pulposus (NP) cells reside in an avascular and hypoxic microenvironment of the intervertebral disc and are predominantly glycolytic due to robust HIF-1 activity. It is generally thought that NP cells contain few functional mitochondria compared with cells that rely on oxidative metabolism. Consequently, the contribution of mitochondria to NP cell metabolism and the role of hypoxia and HIF-1 in mitochondrial homeostasis is poorly understood. Using mitoQC reporter mice, we show for the first time to our knowledge that NP cell mitochondria undergo age-dependent mitophagy in vivo. Mechanistically, in vitro studies suggest that, under hypoxic conditions, mitochondria in primary NP cells undergo HIF-1α-dependent fragmentation, controlled by modulating the levels of key proteins DRP1 and OPA1 that are involved in mitochondrial fission and fusion, respectively. Seahorse assays and steady state metabolic profiling coupled with [1-2-¹³C]-glucose flux analysis revealed that in hypoxia, HIF-1α regulated metabolic flux through coordinating glycolysis and the mitochondrial TCA cycle interactions, thereby controlling the overall biosynthetic capacity of NP cells. We further show that hypoxia and HIF-1α trigger mitophagy in NP cells through the mitochondrial translocation of BNIP3, an inducer of receptor-mediated mitophagy. Surprisingly, however, loss of HIF-1α in vitro and analysis of NP-specific HIF-1α null mice do not show a decrease in mitophagic flux in NP cells but a compensatory increase in NIX and PINK1-Parkin pathways with higher mitochondrial number. Taken together, our studies provide novel mechanistic insights into the complex interplay between hypoxia and HIF-1α signaling on the mitochondrial metabolism and quality control in NP cells. © 2020 American Society for Bone and Mineral Research.     

Growth rate as a prognostic factor in localized invasive cutaneous melanoma

Background Melanoma is a tumour with a very variable progression. Whilst some melanomas grow slowly over many years, others can reach several millimetres in thickness in just a few weeks. Since melanoma is a visible superficial tumour, the information obtained from the clinical interview may be of use to calculate the speed of growth of the melanoma. Objective This study aims to assess the growth rate (GR) of melanomas and the association of this GR with various clinical and pathological factors and their usefulness as prognostic markers for localized invasive cutaneous melanomas. Methods The GR of melanomas was calculated as the ratio of tumour thickness to time of development, as obtained from the clinical history (in millimetres per month). Results Applying the GR calculation to patients with a localized melanoma showed a significant association between melanomas with a GR greater than 0.4 mm per month and an age of 65 years or over, male sex, nodular melanoma, tumour thickness, level of invasion, the presence of ulceration and a high mitotic index. As an independent prognostic factor for overall survival, the GR proved to be significant (P = 0.024). Conclusion The GR of localized cutaneous melanomas may be a possible prognostic factor for survival. Additionally, rapid GR is associated with male patients more advanced in age at diagnosis, which suggests the need to assess new strategies for the early detection of these melanomas.     

Oxygen consumption during sleep in atopic dermatitis

Measurements of oxygen consumption (VO2) were made during sleep in 10 patients with atopic dermatitis. Two groups of healthy children acted as controls. All subjects were studied in bed in an environmental temperature of 24-26 degrees C, and sleep was confirmed during continuous electroencephalographic monitoring. Mean (SD) values of VO2 in sleeping patients who were not scratching ranged from 4.0 (0.4) to 7.4 (0.7), which was not statistically significantly different from control values which ranged from 3.24 (0.3) to 5.56 (0.4). During scratching (while asleep), which occurred in nine out of 10 patients with atopic dermatitis, the mean values of VO2 ranged from 4.5 (0.04) to 10.4 (2.7), and this was significantly higher than the non-scratching patients and the control values. Scratching during sleep in children with atopic dermatitis is associated with increased VO2.     

Organization, expression and polymorphism of the human persyn gene

Persyn is a recently identified member of the synuclein family with a distinct pattern of expression during pre- and postnatal development of the mouse peripheral and central nervous systems. As with other synucleins, persyn is believed to be involved in the pathogenesis of human neurodegenerative diseases. However, in contrast to other synucleins, high levels of persyn mRNA expression were also found in advanced breast carcinomas, suggesting an involvement of the encoded protein in breast tumour progression. Here we have used an antibody specific to human persyn to demonstrate that the level of this protein is increased in ageing cerebral cortex and in breast tumours. We cloned, characterized and sequenced the human persyn genomic locus and localized it to the long arm of chromosome 10 in the q23.2-q23.3 region. Sequence information was used to search for specific mutations in the protein coding regions of persyn mRNA and the persyn gene in breast tumours and tumour cell lines. No tumour-specific mutations were found, but two linked polymorphisms in the coding region were detected, both in mRNA and exons III and IV of the gene. These results suggest that development of breast tumours correlates with overexpression of the wild-type persyn protein. Detailed characterization of the human persyn locus is important for further studies of the involvement of persyn in neurodegeneration and malignancy.      

Duration and predictors of emergency surgical operations - basis for medical management of mass casualty incidents

Background

Hospitals have a critically important role in the management of mass causality incidents (MCI), yet there is little information to assist emergency planners. A significantly limiting factor of a hospital''s capability to treat those affected is its surgical capacity. We therefore intended to provide data about the duration and predictors of life saving operations.

Methods

The data of 20,815 predominantly blunt trauma patients recorded in the Trauma Registry of the German-Trauma-Society was retrospectively analyzed to calculate the duration of life-saving operations as well as their predictors. Inclusion criteria were an ISS ≥ 16 and the performance of relevant ICPM-coded procedures within 6 h of admission.

Results

From 1,228 patients fulfilling the inclusion criteria 1,793 operations could be identified as life-saving operations. Acute injuries to the abdomen accounted for 54.1% followed by head injuries (26.3%), pelvic injuries (11.5%), thoracic injuries (5.0%) and major amputations (3.1%). The mean cut to suture time was 130 min (IQR 65-165 min). Logistic regression revealed 8 variables associated with an emergency operation: AIS of abdomen ≥ 3 (OR 4,00), ISS ≥ 35 (OR 2,94), hemoglobin level ≤ 8 mg/dL (OR 1,40), pulse rate on hospital admission < 40 or > 120/min (OR 1,39), blood pressure on hospital admission < 90 mmHg (OR 1,35), prehospital infusion volume ≥ 2000 ml (OR 1,34), GCS ≤ 8 (OR 1,32) and anisocoria (OR 1,28) on-scene.

Conclusions

The mean operation time of 130 min calculated for emergency life-saving surgical operations provides a realistic guideline for the prospective treatment capacity which can be estimated and projected into an actual incident admission capacity. Knowledge of predictive factors for life-saving emergency operations helps to identify those patients that need most urgent operative treatment in case of blunt MCI.     

Hydrogel-coated textile scaffolds as three-dimensional growth support for human umbilical vein endothelial cells (HUVECs): possibilities as coculture system in liver tissue engineering

Three-dimensional (3-D) scaffolds offer an exciting possibility to develop cocultures of various cell types. Here we report chitosan-collagen hydrogel-coated fabric scaffolds with defined mesh size and fiber diameter for 3-D culture of human umbilical vein endothelial cells (HUVECs). These scaffolds did not require precoating with fibronectin and they supported proper HUVEC attachment and growth. Scaffolds preserved endothelial cell-specific cobblestone morphology and cells were growing in compartments defined by the textile mesh. HUVECs on the scaffold maintained the property of contact inhibition and did not exhibit overgrowth until the end of in vitro culture (day 6). MTT assay showed that cells had preserved mitochondrial functionality. It was also noted that cell number on the chitosan-coated scaffold was lower than that of collagen-coated scaffolds. Calcein AM and ethidium homodimer (EtD-1) dual staining demonstrated presence of viable and metabolically active cells, indicating growth supportive properties of the scaffolds. Actin labeling revealed absence of actin stress fibers and uniform distribution of F-actin in the cells, indicating their proper attachment to the scaffold matrix. Confocal microscopic studies showed that HUVECs growing on the scaffold had preserved functionality as seen by expression of von Willebrand (vW) factor. Observations also revealed that functional HUVECs were growing at various depths in the hydrogel matrix, thus demonstrating the potential of these scaffolds to support 3-D growth of cells. We foresee the application of this scaffold system in the design of liver bioreactors wherein hepatocytes could be cocultured in parallel with endothelial cells to enhance and preserve liver-specific functions.