Monday, March 7, 2022

Understanding orthopedic infections through a different perspective: Microcalorimetry growth curves

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Exp Ther Med. 2022 Apr;23(4):263. doi: 10.3892/etm.2022.11189. Epub 2022 Feb 4.

ABSTRACT

Infectious disease is a serious healthcare problem in orthopedics, as well as other surgical specialties. Accurate and prompt diagnosis, as well as proper care, is critical, as infection of a surgical wound, particularly in the case of arthroplasties or the use of orthopedic implants, can have a catastrophic effect in most cases, necessitating the removal of foreign material. Lyophilized bacteria samples were obtained from Cantacuzino National Institute of Research and Development for Microbiology and Immunology and investigated microcalorimetrically. Isothermal microcalorimetry measures the temperature generated by the multiplication of microorganisms; using an adapted program, it describes the growth curve in real-time according to the received electrical signal. The thermograms of Escherichia coli and Klebsiella pneumonie were anal yzed, and similarities were observed for both the time required for the bacteria to grow and the heat flow generated by their growth. Bacterial microcalorimetry has a variety of benefits, and should be regarded as a means of rapid and accurate diagnosis. Sensitivity is a valuable attribute for a diagnostic technique; when only a few microorganisms are present, microcalorimetric signs of bacterial multiplication can be observed. Microcalorimetry has potential as a simple diagnostic tool in a variety of infections, but further research is needed to ensure that it is used correctly. A thorough investigation (including kinetic analysis) of a reproducible thermal signal of bacterial growth could lead to the creation of new methods for quickly identifying bacteria.

PMID:35251329 | PMC:PMC8892613 | DOI:10.3892/etm.2022.11189

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Role of HSP90α in osteoclast formation and osteoporosis development

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Exp Ther Med. 2022 Apr;23(4):273. doi: 10.3892/etm.2022.11199. Epub 2022 Feb 10.

ABSTRACT

Osteoporosis (OP) is a systemic metabolic bone disease that occurs most frequently in the elderly. The main pathogenesis of OP is excessive proliferation and differentiation of osteoclasts, in which the peroxisome proliferator-activated receptor γ (PPARγ) pathway has a pivotal role. Recently, heat shock protein (HSP)90α has been identified as an important molecular chaperone with PPARγ, which regulates the effect of the PPARγ pathway. The aim of the present study was to investigate the role of HSP90α involved in the regulation of osteoclast formation and the process of osteoporosis. Firstly, the expression of HSP90α in osteoclast differentiation was detected by western blotting in vitro, then the effect of HSP90α inhibition on the formation and differentiation of osteoclasts was examined. Furthermore, the nuclear import of PPARγ wa s also assessed to confirm the synergistic effect of HSP90α. Finally, the inhibitory effect of HSP90α in vivo was explored, using a mouse model of osteoporosis. As a result, in the process of osteoclast differentiation and proliferation, the expression of HSP90α was upregulated. Inhibition of HSP90α could block the formation and differentiation of osteoclasts, and remit osteoporosis in mice. Regarding the underlying mechanism, inhibition of HSP90α could block the nuclear import of PPARγ to inhibit osteoclast differentiation and proliferation. In conclusion, these data indicated that the inhibition of HSP90α could block osteoclast formation and remit osteoporosis by reducing the nuclear import of PPARγ.

PMID:3525133 9 | PMC:PMC8892609 | DOI:10.3892/etm.2022.11199

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Sirtuin 1 participates in intervertebral disc degeneration via the nicotinamide phosphoribosyl transferase/nicotinamide adenine dinucleotide/sirtuin 1 pathway responsible for regulating autophagy of nucleus pulposus cells

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Exp Ther Med. 2022 Apr;23(4):267. doi: 10.3892/etm.2022.11193. Epub 2022 Feb 8.

ABSTRACT

Disc degeneration is the main cause of discogenic low back pain, disc herniation, degenerative stenosis of spinal canal, lumbar spondylolisthesis and other diseases. In the process of intervertebral disc degeneration, water and extracellular matrix of nucleus pulposus tissues are lost, so the normal tension in the intervertebral disc cannot be maintained, which worsens the living environment of nucleus pulposus cells. Low back pain (LBP), with a high incidence rate of disability, has become an increasing health concern and a social and economic problem. The present study aimed to analyze the action mechanisms of nicotinamide phosphoribosyl transferase (Nampt) and sirtuin 1 (SIRT1) in intervertebral disc degeneration (IVDD). In total 26 patients with lumbar disc herniation who had surgical resection at The Third Affiliated Hospital of Jinzhou Medica l University were recruited as the experimental group and their degenerative nucleus pulposus (DNP) tissues of intervertebral disc were collected. In addition, nucleus pulposus tissues of intervertebral disc were collected from 20 patients with burst fracture of lumbar spine at the same hospital (control). Nucleus pulposus cells from primary culturing were separated for subsequent experimentation. LC3 II/I, beclin-1, SIRT1 and NAMPT mRNA and protein expression levels were determined using reverse transcription-quantitative PCR and western blotting, respectively. Nicotinamide adenine dinucleotide (NAD) contents in nucleus pulposus cells was determined by NAD assay kit. The mRNA and protein expression levels of SIRT1 in DNP tissues were reduced compared with the control tissues and decreased with increasing disease severity. The expression of autophagy-associated LC3 II/I and beclin-1 in DNP tissues was reduced compared with control tissues. SIRT1 regulated the LC3 II/I and beclin-1 e xpression levels in nucleus pulposus cells. Treatment with resveratrol and inhibitor of SIRT1 showed that Nampt/NAD+/SIRT1 pathway participated in the process of IVDD by regulating autophagy of nucleus pulposus cells. SIRT1 serves a role in the process of IVDD through Nampt/NAD+/SIRT1 pathway that regulates autophagy of nucleus pulposus cells. SIRT1 may become a biological target for the treatment of IVDD.

PMID:35251333 | PMC:PMC8892612 | DOI:10.3892/etm.2022.11193

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Indigo plant leaf extract inhibits the binding of SARS-CoV-2 spike protein to angiotensin-converting enzyme 2

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Exp Ther Med. 2022 Apr;23(4):274. doi: 10.3892/etm.2022.11200. Epub 2022 Feb 10.

ABSTRACT

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) uses its S1 spike protein to bind to angiotensin-converting enzyme 2 (ACE2) on human cells in the first step of cell entry. Tryptanthrin, extracted from leaves of the indigo plant, Polygonum tinctorium, using d-limonene (17.3 µg/ml), is considered to inhibit ACE2-mediated cell entry of another type of coronavirus, HCoV-NL63. The current study examined whether this extract could inhibit the binding of the SARS-CoV-2 spike protein to ACE2. Binding was quantified as cell-bound fluorescence intensity in live cell cultures in which canine kidney MDCK cells overexpressing ACE2 were incubated with fluorescein-labeled S1 spike protein. When indigo extract, together with S1 protein, was added at 8,650x and 17,300x dilutions, fluorescence intensity decreased in a dose- and S1 extrac t-dependent manner, without affecting cell viability. When 4.0-nM tryptanthrin was added instead of the indigo extract, fluorescence intensity also decreased, but to a lesser degree than with indigo extract. Docking simulation analyses revealed that tryptanthrin readily bound to the receptor-binding domain of the S1 protein, and identified 2- and 7-amino acid sequences as the preferred binding sites. The indigo extract appeared to inhibit S1-ACE2 binding at high dilutions, and evidently contained other inhibitory elements as well as tryptanthrin. This extract may be useful for the prevention or treatment of SARS-CoV-2 infection.

PMID:35251340 | PMC:PMC8892618 | DOI:10.3892/etm.2022.11200

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Inhibition of Sestrin2 overexpression in diabetic cardiomyopathy ameliorates cardiac injury via restoration of mitochondrial function

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Exp Ther Med. 2022 Apr;23(4):265. doi: 10.3892/etm.2022.11191. Epub 2022 Feb 4.

ABSTRACT

Mitochondrial dysfunction-induced apoptosis plays a crucial role in the progression of diabetic cardiomyopathy (DCM). Sestrin2 is an important oxidative stress response protein and is involved in the maintenance of mitochondrial function, especially under stress. The aim of the present study was to investigate the role of Sestrin2 in DCM and to explore the underlying mechanisms. H9c2 cardiomyocytes were induced with high glucose (HG) medium (33 mmol/l glucose) for an in vitro DCM model. C57BL/6 mice were induced for the in vivo DCM model by intraperitoneal streptozotocin injection. H9c2 cardiomyocytes were exposed to HG and infected with lentiviruses to express Sestrin2 short hairpin RNA (shRNA). The study found that cell viability and mitochondrial function were impaired while cell apoptosis and oxidative stress were increased in DCM . Sestrin2 was significantly upregulated in myocardial tissues of DCM mice and H9c2 cardiomyocytes in HG conditions. Downregulation of Sestrin2 increased cell viability, decreased cell apoptosis, and attenuated oxidative stress in H9c2 cells exposed to HG. Moreover, HG-induced mitochondrial injury was alleviated by Sestrin2 silencing. In conclusion, our finding indicated that the inhibition of enhanced Sestrin2 expression ameliorates cardiac injury in DCM, which might be largely attributed to the restoration of mitochondrial function.

PMID:35251331 | PMC:PMC8892608 | DOI:10.3892/etm.2022.11191

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Inhibition of Sestrin2 overexpression in diabetic cardiomyopathy ameliorates cardiac injury via restoration of mitochondrial function

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Exp Ther Med. 2022 Apr;23(4):265. doi: 10.3892/etm.2022.11191. Epub 2022 Feb 4.

ABSTRACT

Mitochondrial dysfunction-induced apoptosis plays a crucial role in the progression of diabetic cardiomyopathy (DCM). Sestrin2 is an important oxidative stress response protein and is involved in the maintenance of mitochondrial function, especially under stress. The aim of the present study was to investigate the role of Sestrin2 in DCM and to explore the underlying mechanisms. H9c2 cardiomyocytes were induced with high glucose (HG) medium (33 mmol/l glucose) for an in vitro DCM model. C57BL/6 mice were induced for the in vivo DCM model by intraperitoneal streptozotocin injection. H9c2 cardiomyocytes were exposed to HG and infected with lentiviruses to express Sestrin2 short hairpin RNA (shRNA). The study found that cell viability and mitochondrial function were impaired while cell apoptosis and oxidative stress were increased in DCM . Sestrin2 was significantly upregulated in myocardial tissues of DCM mice and H9c2 cardiomyocytes in HG conditions. Downregulation of Sestrin2 increased cell viability, decreased cell apoptosis, and attenuated oxidative stress in H9c2 cells exposed to HG. Moreover, HG-induced mitochondrial injury was alleviated by Sestrin2 silencing. In conclusion, our finding indicated that the inhibition of enhanced Sestrin2 expression ameliorates cardiac injury in DCM, which might be largely attributed to the restoration of mitochondrial function.

PMID:35251331 | PMC:PMC8892608 | DOI:10.3892/etm.2022.11191

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Neurological manifestations found in children with multisystem inflammatory syndrome

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Exp Ther Med. 2022 Apr;23(4):261. doi: 10.3892/etm.2022.11187. Epub 2022 Feb 4.

ABSTRACT

The pandemic that resulted from the spread of SARS-CoV-2 viral infections has affected the population worldwide but has characteristically shown a preponderance for affecting adults. However, cases of SARS-CoV-2 infection have been reported in children, showing a systemic echo and severe damage. Multisystem inflammatory syndrome in children (MIS-C) can occur, on average, 4 weeks after the infection of a child with SARS-CoV-2. The aim of the present study was to examine 30 cases of children affected by MIS-C in terms of symptoms, laboratory tests, and evolution. Patients included in the study presented with neurological symptomatology including headache, meningism, and drowsiness. Treatment was administered in concordance with the protocol for MIS-C. The evolution of the patients in the present study was favorable and the symptomatology remitted in days to weeks. The importance of identifying the features of this disease, its treatment, and that the most probable evolution is favorable is significant in the medical world, especially as the pandemic is ongoing.

PMID:35251327 | PMC:PMC8892624 | DOI:10.3892/etm.2022.11187

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Delayed pathologic tibial fracture with chronic osteomyelitis after fibula free flap

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Ear Nose Throat J. 2022 Mar 7:1455613221075775. doi: 10.1177/01455613221075775. Online ahead of print.

ABSTRACT

The reported donor site morbidity of the fibula free flap (FFF) is low; however, several uncommon complications have been reported with tibia fracture rarely being reported. We present a case of a pathological tibial fracture in the setting of chronic osteomyelitis after FFF. A 54-year-old female presented with a benign fibro-osseous lesion of the right mandible an d was treated with mandibulectomy and reconstructed with a left FFF. Approximately 1 year following surgery, the patient presented to the emergency department. Imaging showed a pathological fracture of the distal third of the tibial shaft with persistent erythema and cellulitis of the lateral prior graft harvest site without signs of systemic infection. She was taken to the operating room for irrigation and debridement with culture and biopsy as well as external fixation of the tibial fracture. Intraoperative biopsy and culture demonstrated fracture site change with callus formation and negative culture. The patient was discharged on 6 weeks of IV vancomycin and ceftriaxone. In conclusion, tibial fracture following FFF is an uncommon complication, yet it can be exacerbated by chronic osteomyelitis. This report highlights the importance of close observation and comprehensive wound care of donor sites after free flap harvest for head and neck reconstruction.

PMID:35254923 | DOI:10.1177/01455613221075775

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A case of musculi peronaeus tertius anatomic variation

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Surg Radiol Anat. 2022 Mar 7. doi: 10.1007/s00276-022-02899-7. Online ahead of print.

ABSTRACT

In the routine dissections study for medical postgraduates, a rare anatomical variation between the right leg and the ankle was observed on the Asian male cadaver. The peronaeus tertius muscle in this cadaver's right leg divided into two tendons: the first tendon was attached to the base of the fifth metatarsal bone, and the second tendon was inserted into the base of the fourth me tatarsal bone. The purpose of this paper is to provide detailed anatomical case reports, and to discuss the possible causes and mechanisms of the variation by reviewing relevant literature, so as to provide some reference for future anatomical and clinical related disease research.

PMID:35254492 | DOI:10.1007/s00276-022-02899-7

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Bone fusion in transcele reconstruction of frontoethmoidal meningoencephalocele

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Br J Neurosurg. 2022 Mar 7:1-5. doi: 10.1080/02688697.2022.2047156. Online ahead of print.

ABSTRACT

OBJECTIVE: In surgical correction of frontoethmoidal encephalocele with transcranial approach, advanced facilities are required. While with extracranial approach, though deemed as a safe option in area with limited facilities, procedure was associated with cerebrospinal fluid (CSF) leakage. In this case series, we evaluate the results of transcele reconstruction of frontoethmoidal encephalocele, our approach to reduce the incidence of CSF leaks by focusing on the closure of layers by its embryological derivatives, by its bone fusion.

METHODS: A case series of 14 patients with various types of frontoethmoidal encephalocele who underwent surgery for defect closure using transcele approach between June 2015 and December 2018 was carried out. Surgery was done by a single surgeon in the Department of Neurosurgery of Cipto Mangunkusumo Hospital in Jakarta, Indonesia. We collected the data of intraoperative blood loss and any signs of infection and CSF leak during the patients' one-year follow up. Bone fusion in the defect was evaluated from 3D rendering of head CT scan that was performed before and in 1 year after surgery.

RESULTS: The median percentage of intraoperative blood loss was 5.9% (0.5-18.7%). All 3D rendering of head CT post-surgery during 1 year follow up showed bone fusion and no patient experienced CSF leaks or CNS infections.

CONCLUSIONS: This study showed that using transcele approach in frontoethmoidal reconstruction could give good bone fusion with minimal blood loss and no CSF leaks. We assumed that closure of the layers by its embryological derivative played an important part in bone fusion and in reducing the incidence of CSF leaks, although this finding has to be validated with large-scale studies.

PMID:35254175 | DOI:10.1080/02688697.2022.2047156

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Sunday, March 6, 2022

Influence of Radiation Dose, Photon Energy, and Reconstruction Kernel on rho/z Analysis in Spectral Computer Tomography: A Phantom Study

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In Vivo. 2022 Mar-Apr;36(2):678-686. doi: 10.21873/invivo.12753.

ABSTRACT

BACKGROUND/AIM: The effective atomic number (Zeff) and electron density relative to water (ρe or Rho) of elements can be derived in dual-energy computed tomography (DECT). The aim of this phantom study was to investigate the effect of different photon energies, radiation doses, and reconstruction kernels on Zeff and Rho measured in DECT.

MATERIALS AND METHODS: An anthropomorphic head phantom including five probes of known composition was scanned under three tube-voltage combinations in DECT: Sn140/100 kV, 140/80 kV and Sn140/80 kV with incremented radiation doses. Raw data were reconstructed with four reconstruction kernels (I30, I40, I50, and I70). Rho and Zeff were measured for each probe for all possible combinations of scan and reconstruction parameters.

RESULTS: DECT-based Rho and Zeff close ly approached the reference values with a mean and maximum error of 1.7% and 6.8%, respectively. Rho was lower for 140/80 kV compared with Sn140/100 kV and Sn140/80 kV with differences being 0.009. Zeff differed among all tube voltages with the most prominent difference being 0.28 between 140/80 kV and Sn140/100 kV. Zeff was lower in I70 compared with those of I30 and I40 with a difference of 0.07. Varying radiation dose yielded a variation of 0.0002 in Rho and 0.03 in Z, both considered negligible in practice.

CONCLUSION: DECT comprises a feasible method for the extraction of material-specific information. Slight variations should be taken into account when different radiation doses, photon energies, and kernels are applied; however, they are considered small and in practice not crucial for an effective tissue differentiation.

PMID:35241522 | DOI:10.21873/invivo.12753

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