Identification of Clostridium innocuum hypothetical protein that is cross-reactive with C. difficile anti-toxin antibodies.

OBJECTIVES: Previously considered solely an opportunistic pathogen, Clostridium innocuum (CI) was recently reported in Taiwan to be an emerging cause of antibiotic-associated diarrhea and clinically indistinguishable from Clostridioides difficile (CD) infection. We previously identified CI culture supernatant being cross-reactive with commercial CD toxin enzyme immunoassays. We aimed to identify and characterize the cross-reacting protein and determine whether it functioned as a human toxin. METHODS: We performed western blots using CI culture supernatants and CD anti-toxin antibodies and identified interacting bands. We identified protein(s) using tandem mass spectrometry and evaluated them by cytotoxicity assays. RESULTS: CI, but not CD, was isolated from stool of 12 children and adults with diarrhea. Culture supernatant from 6/12 CI isolates, and an ATCC reference strain, tested positive for CD toxins (total 7/13 isolates) by commercial EIA. Using two of these isolates, we identified two ∼40 kDa hypothetical proteins, CI_01447 and CI_01448, and confirmed cross-reactivity with CD anti-toxin antibodies by enzyme immunoassay and Western blot. Whole-genome sequencing confirmed all 13 isolates contained both genes, which were highly conserved. We observed no cytopathic or cytotoxic effects to HeLa cells when treated with these proteins. We identified amino acid sequence similarity to the NlpC/P60 family of proteins. CONCLUSIONS: Our findings do not suggest CI proteins CI_01448 and CI_01447, which cross-react with antibodies against CD toxins A and B, are toxic to HeLa cells. Further studies are needed to determine the function of these cross-reacting proteins and the potential virulence factors that could be responsible for CI diarrheal disease.

A phase 1 trial of Vorinostat in combination with concurrent chemoradiation therapy in the treatment of advanced staged head and neck squamous cell carcinoma.

Purpose Vorinostat is a potent HDAC inhibitor that sensitizes head and neck squamous cell carcinoma (HNSCC) to cytotoxic therapy while sparing normal epithelium. The primary objective of this Phase I study was to determine the maximally tolerated dose (MTD) and safety of Vorinostat in combination with standard chemoradiation therapy treatment in HNSCC. Patients and Methods Eligible patients had pathologically confirmed Stage III, IVa, IVb HNSCC, that was unresectable or borderline resectable involving the larynx, hypopharynx, nasopharynx, and oropharynx. Vorinostat was administered at the assigned dosage level (100-400 mg, three times weekly) in a standard 3 + 3 dose escalation design. Vorinostat therapy began 1 week prior to initiation of standard, concurrent chemoradiation therapy and continued during the entire course of therapy. Results Twenty six patients met eligibility criteria and completed the entire protocol. The primary tumor sites included tonsil (12), base of tongue (9), posterior pharyngeal wall (1), larynx (4) and hypopharynx (3). Of the 26 patients, 17 were HPV-positive and 9 were HPV-negative. The MTD of Vorinostat was 300 mg administered every other day. Anemia (n = 23/26) and leukopenia (n = 20/26) were the most commonly identified toxicities. The most common Grade3/4 events included leukopenia (n = 11) and lymphopenia (n = 17). No patient had Grade IV mucositis, dermatitis or xerostomia. The median follow time was 33.8 months (range 1.6-82.9 months). Twenty four of 26 (96.2%) patients had a complete response to therapy. Conclusion Vorinostat in combination with concurrent chemoradiation therapy is a safe and highly effective treatment regimen in HNSCC. There was a high rate of complete response to therapy with toxicity rates comparable, if not favorable to existing therapies. Further investigation in Phase II and III trials is strongly recommended.

Enrichment of probiotic ice cream with different dietary fibers: Structural characteristics and culture viability.

This study evaluated the effect of 5 dietary fibers (apple, orange, oat, bamboo, and wheat) on the physicochemical, rheological, and textural characteristics; sensory properties; and culture viability of probiotic ice cream stored at -18°C for 180 d. The presence of orange and apple fibers increased the titratable acidity, decreased the lightness (color) value of the ice creams, and enhanced the red and yellow coloration. Compared with the control sample, the consistency indices and apparent viscosities of the experimental samples increased with the addition of all dietary fibers except oat fiber. The highest viscosity was obtained in the sample fortified with apple fiber, whereas the ice cream containing orange fiber showed the highest hardness after d 60 of storage. The addition of orange and apple fibers significantly increased melting resistance; however, panelists did not generally like these samples in terms of taste-flavor. All ice creams had viable counts of Lactobacillus acidophilus of ≥7 log cfu/g during storage except the samples with orange and bamboo fiber. Bifidobacterium lactis counts were also found to be >6 log cfu/g in those samples until d 150 of storage.

Serum endocan level and the severity of spinal cord injury.

OBJECTIVE: To determine whether the endocan level may be related to the severity of spinal cord injury. BACKGROUND: Several biomarkers were evaluated for this purpose, but endocan has never been studied before. It is correlated to endothelial dysfunction and ischemia, which are the characteristics of spinal cord injury in most cases. MATERIAL AND METHODS: A total of 21 male Sprague-Dawley rats weighing 300-350 g were randomly divided into three groups. In Group I, only a laminectomy was performed; in Group II, a mild SCI was performed after laminectomy; and, in Group III, a severe SCI was performed after laminectomy. At 48 h after the injury, after neurological assessment by Tarlov method, all animals were euthanized. A 5 cc blood sample was drawn for biochemical analysis, and spinal cord tissues were removed for histopathological examination. RESULTS: The difference between Groups I and III was statistically significant (p < 0.05). There was also a moderately positive correlation between the severity of SCI and the endocan level (r = 0.59, p < 0.05). CONCLUSION: The endocan level may be used as an indicator to determine prognosis after SCI (Tab. 1, Fig. 2, Ref. 24). Text in PDF www.elis.sk.

Follow up of a family with asymptomatic compound long QT syndrome mutations.

Long QT syndromes (LQTS) are a cause of syncope and sudden death and present as a long QT interval on the surface ECG. The mortality of the condition may be quite variable among affected individuals. Hundreds of mutations in more than ten genes are identified as responsible for almost all patients with LQTS. Compound mutations are reported in different series as 4.5 and 7.9% and are associated with poor outcome. Beta blockers are the mainstay of therapy. The use of intracardiac defibrillators (ICD) is widely considered in patients at high risk for sudden death. Herein, we report a case of LQTS with compound mutations of KCNQ1 and SCN5a. Although ICD implantation was advised due to high cardiac event risk, the patient followed with beta blocker treatment for 15 years without any syncope or palpitations.

Vaccination is preventing development of placental pathologies in SARS-CoV-2 infected pregnant patients.

INTRODUCTION: The coronavirus disease (COVID-19) has created a serious health problem in pregnant people. We aimed to address whether vaccination can prevent development of placental disease in SARS-CoV-2 infected mothers. METHODS: We reported the pathology findings obtained from routine histopathological examination of placentas of overall 38 cases. RESULTS: We found low prevalence of placental pathology in vaccinated pregnant people with active SARS-CoV-2 infection in comparison to those unvaccinated cases. CONCLUSION: Based on our findings, SARS-CoV-2 vaccination can prevent development of placental pathological lesions and may lower the risk of serious illness in pregnant people.

Anterolateral thigh butterfly free flap for reconstruction of laryngopharyngeal defect: surgical considerations.

BACKGROUND: Reconstruction of a pharyngoesophageal defect remains a challenging problem, especially with involvement of the neck skin. This study aimed to demonstrate the surgical technique of utilising a butterfly modification of the anterolateral thigh flap. RESULTS: Reconstruction of the pharyngoesophageal defect was accomplished using the butterfly modification of the anterolateral thigh free flap. The flap was tubed on the leg while still being attached to the pedicle, to minimise the ischaemia time. CONCLUSION: Butterfly anterolateral thigh free flap allows for multi-layer closure of the neopharynx and can be utilised for reconstruction of pharyngoesophageal and neck skin defects.

High dose zoledronic acid increases ischemia-reperfusion damage of the liver.

OBJECTIVE: Zoledronic acid (ZA), a nitrogen-containing bisphosphonate, has been reported to exhibit a protective effect against cancers and prevent bone fractures. It also induces apoptosis by increasing proinflammatory cytokines and oxidative stress. Oxidative stress increases significantly during ischemia-reperfusion (IR) injury. The liver is highly sensitive to IR injury. In this study, we aim to investigate whether high-dose ZA treatment affects the liver during IR. MATERIALS AND METHODS: We used twenty-one Sprague-Dawley male rats in our study, and they were subdivided randomly into three groups, each containing seven rats. A single dose of 100 µg/kg ZA was administered via the intraperitoneal route in the ZA group. Forty-eight hours after the ZA administration, infrarenal abdominal aortic cross ligation was performed on the ZA and IR groups. After 2 hours of ischemia, 2 hours of reperfusion was applied. RESULTS: The malondialdehyde (MDA) level of the control group was significantly lower than the IR (p = 0.006) and ZA (p<0.001) groups. However, the superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) values of the control group were significantly higher than the values of the IR group (p<0.05, p<0.001, and p<0.05) and ZA group (p = 0.002, p<0.001, and p<0.001). Caspase-3 activity was significantly higher in the IR group as compared to the control group (p<0.001). The caspase-3 activity in the ZA group, on the other hand, was higher than both the control (p<0.001) and IR groups (p<0.001). CONCLUSIONS: High-dose ZA may exacerbate liver injury during IR by increasing reactive oxygen species production and apoptosis.

A novel presenilin1 mutation (Q223R) associated with early onset Alzheimer's disease, dysarthria and spastic paraparesis and decreased Abeta levels in CSF.

BACKGROUND AND PURPOSE: A novel presenilin1 (PSEN1) mutation associated with dementia and spastic paraplegia in a family with five affected individuals is described. The index patient was a 35-year-old man presenting with cognitive decline, behavioural symptoms, dysarthria, and gait disorder due to spasticity. METHODS AND RESULTS: Genetic analysis revealed a missense mutation Gln223Arg in exon 7. Initial CSF analysis revealed drastically decreased Abeta42 level despite marginally decreased FDG metabolism. CONCLUSION: Cerebrospinal fluid biomarker analysis might point towards genetic analysis of PSEN1 in patients with positive family history and age of onset below 60 years.

Serum vitamin D status and bone mineral density in fibromyalgia.

OBJECTIVES: To compare serum vitamin D levels and bone mineral density (BMD) values in patients with fibromyalgia and healthy controls. BACKGROUND: The so far available reports of low levels of vitamin D and low BMD values in patients with fibromyalgia are inconsistent. METHODS: Serum 25-hydroxyvitamin D (25-OHD) levels and BMD values were measured in thirty women with fibromyalgia and compared with thirty age-matched healthy women. Serum calcium, phosphorus, alkaline phosphatase and parathyroid hormone (PTH) levels were also measured. All participants completed the Fibromyalgia Impact Questionnaire (FIQ) and Hospital Anxiety and Depression Score (HADS). Pain severity was assessed with visual analog scale (VAS). RESULTS: Mean serum 25-OHD levels did not differ between the groups (fibromyalgia 10.57 +/- 10.46, controls 10.87 +/- 5.52 ng/l; p=0.89); nor did the frequency of vitamin D deficiency (25-OHD < or = 20 ng/l) in each group (fibromyalgia 86.7%, controls 96.7%; p=0.353). Although, mean serum PTH level was found significantly higher in fibromyalgic patients than in controls (p=0.014), only one patient and two of controls had barely elevated PTH levels. There was no relationship between vitamin D level and FIQ score (p=0.707) or HADS (p=0.824) or pain VAS (p=0.414). BMD values in the patients with fibromyalgia were comparable to those in controls at both, the lumbar spine (p=0.866) and femur neck (p=0.61). CONCLUSION: Neither vitamin D levels nor BMD values are different between women with and without fibromyalgia. In this cross-sectional study, mean serum PTH level was found higher in the fibromyalgic patients than in controls. Nevertheless, in order to confirm the findings of this preliminary study it is still necessary to perform a controlled longitudinal study (Tab. 2, Fig. 2, Ref. 35). Full Text in free PDF www.bmj.sk.

Complete Genome Sequence of Clostridium innocuum Strain ATCC 14501.

We report the complete genome sequence of Clostridium innocuum ATCC 14501, which was isolated in 1962 from an appendiceal abscess. At that time, the isolated strain was designated C. innocuum, given its suspected lack of virulence, but recent reports suggest that C. innocuum is an emerging pathogen.

Complete Genome Sequence of Clostridium innocuum Strain LC-LUMC-CI-001, Isolated from a Patient with Recurrent Antibiotic-Associated Diarrhea.

Here, we report the complete genome sequence of Clostridium innocuum strain LC-LUMC-CI-001. As recently as 2018, C. innocuum was generally considered a benign gastrointestinal microorganism. This strain was isolated from the stool of a patient with recurrent Clostridioides difficile infection-like illnesses.

Which agent should be used to reduce ischemia-reperfusion injury after testicular torsion: a comparative animal experiment.

INTRODUCTION: Testicular torsion is an acute urological emergency that causes severe damage of testis. In order to prevent testicular damage, early diagnosis and surgical intervention is essential. Also pain management in both pre-operative and postoperative periods remains a challenging entity. OBJECTIVE: In this study, we aimed to determine the possible positive effects of three different analgesics (ibuprofen, metamizole, and paracetamol), which are widely used in clinical practice on testicular tissue, in addition to pain control. STUDY DESIGN: Forty prepubertal rats (180-210 g) were divided into five experimental groups. Group 1 was sham group in which the left testis was brought out through a scrotal incision and then replaced in the scrotum without torsion. Group 2 was control group (only 0.9% NaCl was applied). Also in group 3, paracetamol, in group 4, ibuprofen, and in group 5, metamizole sodium was applied 1 h after the torsion. Torsion duration was planned as 4 h for all groups. RESULTS: In the biochemical evaluation, malondialdehyde (MDA), myeloperoxidase (MPO), and total nitrate (NO) levels were measured in the testicular tissue. All groups were compared with group 2 (control group). In group 3, although the MDA level was lower and the MPO level was found to be higher, these were not statistically significant. In group 4, the NO level was low but statistically significant. Histological findings were evaluated due to Cosentino's classification, and the scores of group 4 were better than all groups. DISCUSSION: In this study, severe damage was observed at the end of torsion period of 4 h. This is in line with previous published data. The beneficial effects of all three drugs have been observed. CONCLUSIONS: Biochemical results did not clearly highlight any agents. According to pathology results, metamizole was better than paracetamol and the most ideal analgesic preparation was observed as ibuprofen.

Acro-cardio-facial syndrome associated with neuroepithelial cyst: a case report.

Acro-cardio-facial syndrome (ACFS) is a very rare genetic syndrome. Only 5 patients have been reported in the literature so far. A female neonate presented with limb abnormalities, cleft palate and congenital heart disease was diagnosed as ACFS. Her cranial magnetic resonance imaging revealed a huge cerebral neuroepithelial cyst. To our knowledge, this is the first case of ACFS in the literature associated with a neuroepithelial cyst in the brain.

Sympathetic skin responses in patients with hyperthyroidism.

BACKGROUND: The aim of this study was to investigate the disorders of sympathetic nervous system in patients with hyperthyroidism using sympathetic skin response (SSR). MATERIAL AND METHODS: Twenty-two newly diagnosed cases with hyperthyroidism were included in the study. The results were compared with those of 20 healthy controls. SSR was recorded with the contralateral electrical stimulation of the median nerve (of the upper extremities) and tibial nerve (of the lower extremities) with active electrodes placed on palms and soles and reference electrodes attached on the dorsal aspects of hands and feet. RESULTS: Ages of the cases with hyperthyroidism and controls ranged between 15-65 years (mean: 46.7 +/- 15.0 years) and 24-62 years (mean: 39.6 +/- 9.8 years) respectively (p > 0.05). In all the control subjects SSR could be obtained, while from the lower extremities of 4 cases with hyperthyroidism (18.0%) SSR could not be elicited. Mean SSR latencies of lower extremities were found significantly longer than control group (p < 0. 05). No difference was detected between mean amplitudes of SSR in upper and lower extremities. CONCLUSION: These findings suggest that SSR is useful for investigation of sympathetic nervous system involvement in cases with hyperthyroidism.

Inhibition of vascular endothelial growth factor-induced retinal neovascularization by retinoic acid in experimental retinopathy of prematurity.

Vascular endothelial growth factor (VEGF) has an important role in the pathogenesis of retinopathy of prematurity (ROP) and inhibition of VEGF expression in the neovascular phase might prevent destructive neovascularization in ROP. It is suggested that retinoids exert a highly potent antiangiogenic activity by inhibiting VEGF expression. The aim of this study was to demonstrate the preventive effect of retinoic acid (RA) on the VEGF-induced retinal neovascularization in a rat model of ROP. Wistar albino rats were placed into incubators at birth and exposed to an atmosphere alternating between 50 % and 10 % O(2) every 24 hours. After 14 days, the animals were removed to room air and received either an intraperitoneal injection of RA (5 mg/kg/day) (n=9) or saline (n=4) daily for six days, and sacrificed at 21 days. Other rats (n=4) were raised in room air and served as age-matched controls. The globe of each eye was cut through the cornea and embedded in paraffin. Serial sections were stained with hematoxylin-eosin for quantification of neovascular nuclei. The avidin-biotin peroxidase method was performed for evaluation of VEGF expression. The average number of neovascular nuclei was significantly lower in the control group compared to that in the ROP groups. In addition, it significantly decreased in the RA-treated ROP group compared to that of the saline-administrated ROP group. VEGF immunostaining was overall negative in room air-exposed rats. The VEGF immunostaining score significantly decreased in the RA-treated ROP group compared to that in the saline-administered ROP group. RA treatment might be beneficial in preventing neovascularization resulting from oxygen-induced retinopathy by downregulation of VEGF expression.

Erythropoietin attenuates lipopolysaccharide-induced splenic and thymic apoptosis in rats.

Apoptosis of lymphoid tissues during sepsis is well documented and linked to the pathobiology of organ failure and death. In this study, we evaluated the effect of a single dose of recombinant erythropoietin (EPO) on thymic and splenic apoptosis in an endotoxic sepsis model. Young male Wistar rats were divided into 3 groups and administered intraperitoneally (IP) either normal saline; lipopolysaccharide (LPS) 10 mg/kg; or EPO (5000 U/kg) 30 min before lipopolysaccharide. Six hours following LPS administration animals were sacrificed. Apoptosis was assessed by hematoxylin-eosin staining, terminal deoxynucleotide transferase-mediated fluorescein-dUTP nick end labeling (TUNEL), and caspase-3 immunostaining. When compared with animals given LPS, animals pretreated with EPO displayed reduced splenic and thymic TUNEL positivity of 44+/-3 (p<0.05) and 143+/-4 (p<0.05) nuclei per high power field (hpf), respectively. Caspase-3 positivity was also significantly reduced in the spleen and thymus, with 31+/-4 (p<0.05) and 93+/-3 (p<0.05) positive stained nuclei per hpf, respectively. Serum nitrite levels were elevated in animals given lipopolysaccharide. Pretreatment with EPO attenuated the increase in nitrite levels; however, this did not reach statistical significance. We conclude that a single dose of recombinant erythropoietin can reduce thymic and splenic apoptosis associated with lipopolysaccharide administration.