Effect of Helicobacter pylori Vacuolating Toxin on Maturation and Extracellular Release of Procathepsin D and on Epidermal Growth Factor Degradation

Abstract The effect of vacuolating toxin (VacA) fromHelicobacter pylori on endosomal and lysosomal functions was studied by following procathepsin D maturation and epidermal growth factor (EGF) degradation in HeLa cells exposed to the toxin. VacA inhibited the conversion of procathepsin D (53 kDa) into both the intermediate (47 kDa) and the mature (31 kDa) form. Nonprocessed cathepsin D was partly retained inside cells and partly secreted in the extracellular medium via the constitutive secretion pathway. Intracellular degradation of EGF was also inhibited by VacA with a similar dose-response curve. VacA did not alter endocytosis, cell surface recycling, and retrograde transport from plasma membrane totrans-Golgi network and endoplasmic reticulum, as estimated by using transferrin, diphtheria toxin, and ricin as tracers. Subcellular fractionation of intoxicated cells showed that procathepsin D and nondegraded EGF accumulate in lysosomes. Measurements of intracellular acidification with fluorescein isothiocyanate-dextran revealed a partial neutralization of the lumen of endosomes and lysosomes, sufficient to account for both mistargeting of procathepsin D outside the cell and the decreased activity of lysosomal proteases

The myloglossus muscle: anatomical and clinical observations

The myloglossus muscle is considered an anomalous muscle among the extrinsic muscle of the tongue. In the past, only few Authors provided an anatomical description of the myloglossus muscle (Valenti, 1925; Jude, 1973; Gruber, 1980) and recently a description of myloglossus muscle in Japanese cadaver was reported (Nakajima and Nakamura, 2008). Dissection studies showed that the myloglossus muscle arises from the inner surface of the mandible between the alveolar process and the distal part of the mylohyoid groove and inserts into the tongue root, joining the palatoglossus muscle. Some anatomical variations regarding its origin could be present: it could originate from stylomandibular ligament or partially replace the styloglossus muscle. In patients, with discrete muscle trophism, the myloglossus muscle provides a lateral to medial mucosal fold in the posterior portion of the sublingual sulcus that is evident when the tongue is controlaterally moved. On the contrary, in patients with poor muscle trophism these mucosal folds were not observed. The myloglossus muscle acts primarily as an antagonist of both the controlateral muscle and other muscles that move the tongue toward the opposite side; moreover, it acts together with the controlateral and the palatoglossus muscle, determining the upward movements of the tongue and pharynx, especially in the last phase of deglutition. Its location should be considered to well determine the distal lingual extension of the removable prosthesis. Gruber W. 1880. Uber den musculus myloglossus bei mangel und vorkommen des styloglossus. Arch Path Anat Physiol Klin Med 81:453-457. Jude HD. 1973. Anatomiche untersuchungen uber extensionmoglichkeiten unterer total prothesen im retromolaren raum. Dtsch Zahnarztl Z 28:486. Nakajima K, Nakamura M. Rare case of myloglossus in Japanese cadaver: anatomical and developmental considerations. Anat Sci Int. 2008; 83: 1-5 Valenti G. 1925. Sur un muscle mandibulo-glosse (M. Mylo-Glossus Wood). Arch Ital Biol 75: 77

Inhibition of ERK5 elicits cellular senescence in melanoma via the cyclin-dependent kinase inhibitor p21

2021 The Authors.Melanoma is the deadliest skin cancer with a very poor prognosis in advanced stages. Although targeted and immune therapies have improved survival, not all patients benefit from these treatments. The mitogen-activated protein kinase ERK5 supports the growth of melanoma cells in vitro and in vivo. However, ERK5 inhibition results in cell-cycle arrest rather than appreciable apoptosis. To clarify the role of ERK5 in melanoma growth, we performed transcriptomic analyses following ERK5 knockdown in melanoma cells expressing BRAFV600E and found that cellular senescence was among the most affected processes. In melanoma cells expressing either wild-type or mutant (V600E) BRAF, both genetic and pharmacologic inhibition of ERK5 elicited cellular senescence, as observed by a marked increase in senescence-associated β-galactosidase activity and p21 expression. In addition, depletion of ERK5 from melanoma cells resulted in increased levels of CXCL1, CXCL8, and CCL20, proteins typically involved in the senescence-associated secretory phenotype. Knockdown of p21 suppressed the induction of cellular senescence by ERK5 blockade, pointing to p21 as a key mediator of this process. In vivo, ERK5 knockdown or inhibition with XMD8–92 in melanoma xenografts promoted cellular senescence. Based on these results, small-molecule compounds targeting ERK5 constitute a rational series of prosenescence drugs that may be exploited for melanoma treatment.The work in E. Rovida’s lab was supported by grants from Associazione Italiana per la Ricerca sul Cancro (AIRC, IG-15282 and IG-21349), by Ente Fondazione Cassa di Risparmio di Firenze (ECRF), and Universita degli Studi di Firenze (Fondo di Ateneo ex-60%). A. Tubita was supported by a “Carlo Zanotti” Fondazione Italiana per la Ricerca sul Cancro (FIRC)-AIRC fellowship (ID-23847)

Erratum: Antibodies from multiple sclerosis patients preferentially recognize hyperglucosylated adhesin of non-typeable Haemophilus influenzae

Scientific Reports 6: Article number: 39430; published online: 23 December 2016; updated: 22 March 2017. The original version of this Article contained typographical errors in the Abstract. ‘In autoimmune diseases, there have been proposals that exogenous “molecular triggers”, i.e., specific this should be ‘non-self antigens’ accompanying infectious agents, might disrupt control of the adaptive immune system resulting in serious pathologies’.</jats:p

Atlas rotation and mandibular deviation by Cone Beam CT

Cervical vertebrae and mandible are functionally related and some evidences suggest a strong correlation between their relative position and orientation (Huggare et al., 1996; Nisayif et al., 2005). In this study TC Dental Scan with cone beam technology was used to study the relationship between atlas and mandibular rotation in 205 patients. Using a digitalized images analyser, we calculated the axial rotation of atlas and mandible, measuring the angle with respect to the frontal plane. We found that 80.98% of patients presented the axial rotation of the mandible in the same direction of atlas rotation compared with 19.02% of patients that presented opposite directions. Among the consistent group, 71.08% of patients had a left rotation compared with 28.92% that had a right rotation. Moreover, considering the absolute values of the rotation, we observed that the atlas had a more marked rotation with respect to the mandible and that the values of left rotations were higher with respect to the value recorded for right rotations both for the mandible and atlas measurements. This study represents a starting point to better characterize the relationship between atlas and mandible; further studies are necessary to better understand the importance of this data from a functional and clinical point of view

Antibodies from multiple sclerosis patients preferentially recognize hyperglucosylated adhesin of non-typeable Haemophilus influenzae

In autoimmune diseases, there have been proposals that exogenous "molecular triggers", i.e., specific this should be 'non-self antigens' accompanying infectious agents, might disrupt control of the adaptive immune system resulting in serious pathologies. The etiology of the multiple sclerosis (MS) remains unclear. However, epidemiologic data suggest that exposure to infectious agents may be associated with increased MS risk and progression may be linked to exogenous, bacterially-derived, antigenic molecules, mimicking mammalian cell surface glycoconjugates triggering autoimmune responses. Previously, antibodies specific to a gluco-asparagine (N-Glc) glycopeptide, CSF114(N-Glc), were identified in sera of an MS patient subpopulation. Since the human glycoproteome repertoire lacks this uniquely modified amino acid, we turned our attention to bacteria, i.e., Haemophilus influenzae, expressing cell-surface adhesins including N-Glc, to establish a connection between H. influenzae infection and MS. We exploited the biosynthetic machinery from the opportunistic pathogen H. influenzae (and the homologous enzymes from A. pleuropneumoniae) to produce a unique set of defined glucosylated adhesin proteins. Interestingly we revealed that a hyperglucosylated protein domain, based on the cell-surface adhesin HMW1A, is preferentially recognized by antibodies from sera of an MS patient subpopulation. In conclusion the hyperglucosylated adhesin is the first example of an N-glucosylated native antigen that can be considered a relevant candidate for triggering pathogenic antibodies in MS

Morphometric analysis of Huguier’s canal by Cone Beam CT

The middle ear and the stomatognatic system are closely anatomically and functionally related. The anterior chordal canal of Huguier connects the temporomandibular joint (TMJ) and the middle ear. This canal is formed by the inferior process of tegment tympani and the sphenoid bone and it is located at the medial end of the petrotympanic fissure. To data, few studies aimed to describe Huguier’s canal morphology and its related structures (Toth et al., 2006; Sato et al., 2008; Aristeguieta et al., 2009). The aim of this study is to describe the radiological anatomy of the Huguier’s canal using cone beam CT (CBCT, Scanora 3D, Soredex). We measured 438 Huguier’s canals from 219 human skulls (Section of Anthropology and Ethnology, Museum of Natural History, Florence, Italy). The measurements were made at three levels: 1) near the TMJ (lateral-glenoidal side) that was 1.961 ± 0.472 mm; 2) the narrowest point of the middle area that was 0.494 ± 0.24 mm; 3) near the middle ear (medial, acoustic meatus side) that was 1.085 ± 0.354 mm. 21 on 439 Huguier’s canal (4.79%) were ossificated: 1 only in the medial side, 11 only in the middle area and 9 in all the three levels. Considering the high number of measurements, the values obtained were comparable, suggesting that CBCT can be useful to detect these anatomical details

Current Knowledge on Endocrine Disrupting Chemicals (EDCs) from Animal Biology to Humans, from Pregnancy to Adulthood: Highlights from a National Italian Meeting

Wildlife has often presented and suggested the effects of endocrine disrupting chemicals (EDCs). Animal studies have given us an important opportunity to understand the mechanisms of action of many chemicals on the endocrine system and on neurodevelopment and behaviour, and to evaluate the effects of doses, time and duration of exposure. Although results are sometimes conflicting because of confounding factors, epidemiological studies in humans suggest effects of EDCs on prenatal growth, thyroid function, glucose metabolism and obesity, puberty, fertility, and on carcinogenesis mainly through epigenetic mechanisms. This manuscript reviews the reports of a multidisciplinary national meeting on this topic

Canagliflozin and renal outcomes in type 2 diabetes and nephropathy

BACKGROUND Type 2 diabetes mellitus is the leading cause of kidney failure worldwide, but few effective long-term treatments are available. In cardiovascular trials of inhibitors of sodium–glucose cotransporter 2 (SGLT2), exploratory results have suggested that such drugs may improve renal outcomes in patients with type 2 diabetes. METHODS In this double-blind, randomized trial, we assigned patients with type 2 diabetes and albuminuric chronic kidney disease to receive canagliflozin, an oral SGLT2 inhibitor, at a dose of 100 mg daily or placebo. All the patients had an estimated glomerular filtration rate (GFR) of 30 to &lt;90 ml per minute per 1.73 m2 of body-surface area and albuminuria (ratio of albumin [mg] to creatinine [g], &gt;300 to 5000) and were treated with renin–angiotensin system blockade. The primary outcome was a composite of end-stage kidney disease (dialysis, transplantation, or a sustained estimated GFR of &lt;15 ml per minute per 1.73 m2), a doubling of the serum creatinine level, or death from renal or cardiovascular causes. Prespecified secondary outcomes were tested hierarchically. RESULTS The trial was stopped early after a planned interim analysis on the recommendation of the data and safety monitoring committee. At that time, 4401 patients had undergone randomization, with a median follow-up of 2.62 years. The relative risk of the primary outcome was 30% lower in the canagliflozin group than in the placebo group, with event rates of 43.2 and 61.2 per 1000 patient-years, respectively (hazard ratio, 0.70; 95% confidence interval [CI], 0.59 to 0.82; P=0.00001). The relative risk of the renal-specific composite of end-stage kidney disease, a doubling of the creatinine level, or death from renal causes was lower by 34% (hazard ratio, 0.66; 95% CI, 0.53 to 0.81; P&lt;0.001), and the relative risk of end-stage kidney disease was lower by 32% (hazard ratio, 0.68; 95% CI, 0.54 to 0.86; P=0.002). The canagliflozin group also had a lower risk of cardiovascular death, myocardial infarction, or stroke (hazard ratio, 0.80; 95% CI, 0.67 to 0.95; P=0.01) and hospitalization for heart failure (hazard ratio, 0.61; 95% CI, 0.47 to 0.80; P&lt;0.001). There were no significant differences in rates of amputation or fracture. CONCLUSIONS In patients with type 2 diabetes and kidney disease, the risk of kidney failure and cardiovascular events was lower in the canagliflozin group than in the placebo group at a median follow-up of 2.62 years