Gene expression profiles of antigenic proteins of third stage larvae of the zoonotic nematode Anisakis pegreffii in response to temperature conditions

Anisakis pegreffii, a recognised etiological agent of human anisakiasis, is a parasite of homeothermic hosts at the adult stage and of ectothermic hosts at the third larval stage. Among distinct factors, temperature appears to be crucial in affecting parasite hatching, moulting and to modulate parasite-host interaction. In the present study, we investigated the gene transcripts of proteins having an antigenic role among excretory secretory products (ESPs) (i.e., a Kunitz-type trypsin inhibitor, A.peg-1; a glycoprotein, A.peg-7; and the myoglobin, A.peg-13) after 24 h, in A. pegreffii larvae maintained in vitro, under controlled temperature conditions. Temperatures were 37 °C and 20 °C, resembling respectively homeothermic and ectothermic hosts conditions, and 7 °C, the cold stress condition post mortem of the fish host. Primers of genes coding for these ESPs to be used in quantitative real-time PCR were newly designed, and qRT-PCR conditions developed. Expression profiles of the genes A.peg-1 and A.peg-13 were significantly up-regulated at 20 °C and 37 °C, with respect to the control (larvae kept at 2 °C for 24 h). Conversely, transcript profiles of A.peg-7 did not significantly change among the chosen temperature conditions. In accordance with the observed transcript profiles, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed the presence of the three target ESPs at 37 °C, while only A.peg-13 was observed at 7 °C. The results suggest that temperature conditions do regulate the gene expression profiles of A.peg-1 and A.peg-13 in A. pegreffii larvae. However, regulation of the glycoprotein A.peg-7 is likely to be related to other factors such as the host's immune response

Species-specific Real Time-PCR primers/probe systems to identify fish parasites of the genera Anisakis, Pseudoterranova and Hysterothylacium (Nematoda: Ascaridoidea)

Ascaridoid nematodes belonging to the genera Anisakis and Pseudoterranova are heteroxenous parasites, involving marine mammals as definitive hosts in their life-cycles, whereas crustaceans (krill), fish and squids acting as intermediate/paratenic hosts. These parasites are considered among the most important biological hazards present in “seafood” products. Indeed, larval stages of the Anisakis and Pseudoterranova have been reported as etiological agents of human infections (anisakidosis). We developed a primers/probe system for the identification of five species of anisakid nematodes belonging to the genera Anisakis (i.e. A. pegreffii and A. simplex (s. s.)), and Pseudoterranova (i.e. P. decipiens (s. s.), P. krabbei and P. bulbosa) to be used in a real time polymerase chain reaction (RT-PCR) with specific primers based on the mtDNA cox2 gene. Because those anisakid species could be also found in co-infection in some fish species with the raphidascarid nematode Hysterothylacium aduncum, a species-specific primer probe system to be used in RT-PCR for this nematode species was also developed. The detection limit and specificity of the primer/probe systems were evaluated for each of the six nematode species. Singleplex and multiplex RT-PCR protocols were defined and tested. The detection limit of the nematode species tissue was lower than 0.0006 ng/μl. Efficiency (E) of primers/probe systems developed was carried out by standard curve; E value varied between 2.015 and 2.11, with respect to a perfect reaction efficiency value of E = 2. Considering the sensibility and quantitative nature of the assays, the new primers/probe system may represent a useful tool for future basic and applied research that focuses on the identification of Anisakis spp., Pseudoterranova spp. and H. aduncum larvae in fish, even in co-infections, with a potential for application in fish farming, fish processing industries, fish markets, and food producers

Predictive factors of hospitalization related to the caregiver burden in older adults presenting to the emergency department

Background Long-term care for the elderly by their family members represents a serious burden in Italy. The physical and psychological health of informal caregivers is a growing public health issue. Old patients often seek urgent medical attention in the Emergency Department (ED) and hospitalisation is frequent event among the elderly. Aim Aim of the study was (1) to investigate the burden of care among the caregivers of old patients; (2) to examine the influence of the burden experienced by the caregivers on ED and hospital admissions of the elderly. Methods We conducted a descriptive study of patients aged 75 years or older and their caregiver admitted to the ED from 10/1/15 to 6/10/15 (77 patient-caregiver pairs). The caregivers were evaluated using the Caregiver Burden Inventory (CBI). A case manager collected the patient's data. Results CBI score is the highest among patients seeking ED evaluation due to caregiver's concern. The majority of the elderly admitted to the ED whose caregiver shows elevated emotional burden at the CBI do not present with serious or urgent medical condition and are not hospitalised. Emotional burden is the highest among the caregivers of demented subjects who share the same house. Conclusion Our findings indicate that the burden experienced by caregiving family members plays a role in elderly people avoidable ED visits

Psychiatric referral in the terminally ill patient

The distinction between physiological and dysfunctional emotions in end-of-life care may be hard, for a twofold reason: on the one hand, the patient as a subject, with specific clinical features, personality, system of values; on the other hand, the clinical judgment by involved health professionals, particularly their specific cut-offs in discrimination between normal suffering and psychopathology. Both excessive/untimely medicalization and underestimation of medical conditions such as anxiety, depression, suicidal ideation, and insomnia may be a risk while dealing with end-of-life patients. Prompt, reliable psychiatric diagnosing contributes significantly to the major goal of dignity in death. The aims of a psychiatric consultation for patients with end-stage diseases should be: controlling concomitant psychiatric symptoms, managing pain and physical symptoms, assisting patient and relatives in the crisis-management, mediating conflicts between patient, family and ward personnel, and planning advocacy

Clinical features of colorectal cancer patients in advanced age: a population-based approach

In the immediate future, the number of geriatric patients will continue to rise; consequently we should expect an increase of colorectal cancer, a disease of the elderly population. Through the data of a Cancer Registry, we examined (a) the effect of ageing on the main features of colorectal cancer; (b) changes in management, especially for individuals older than 80 years; and (c) changes in prognosis and survival in subgroups of patients with different age. The Registry provided information on colorectal cancer up to 2010 (27 years). A total of 5293 patients were registered; these were divided into three groups: A (0â64 years), B (65â79) and C (80 or more). Three periods of observation were chosen: 1 (1984â1992), 2 (1993â2001) and 3 (2001â2010). Group A included 1571 patients (29 %), Group B 2539 (48 %) and Group C 1183 (22.3 %). The fraction of old individuals increased during the 27 years of the investigation. In these patients, tumours were predominantly localized to the right colon (42.6 %). The rate of surgery and ratio between curative and palliative approaches were similar among the three groups (pÂ&nbsp

Clinical staging systems for hepatocellular carcinoma: Comparison with estrogen receptor classification

Clinical staging systems for hepatocellular carcinoma: Comparison with estrogen receptor classificatio