The development of ultrasensitive immunological methods for the detection of protein polymorphisms

This thesis describes the development of ultrasensitive methods for the immunological detection of two highly polymorphic human proteins, group-specific component (GC) and phosphoglucomutase-1 (PGM) following separation by high resolution isoelectric focusing (IEF) and transfer to immobilising membranes. All the steps involved in the immunoblotting procedure were investigated and the procedure optimised with a view to its application in the field of Forensic Science. In the case of the GC polymorphism, a 3000 fold increase in the sensitivity of detection was achieved compared with immunofixation methods. The improved immunoblotting procedure allowed the phenotyping of GC from forensic bloodstains, semen stains and post-coital vaginal swabs. A high titre sheep anti-rabbit muscle PGM antiserum was produced that was capable of detecting both rabbit and human PGMl isozymes on immunoblots, following sodium-dodecyl-sulphate gel electrophoresis or IEF, of tissues expressing high levels of the enzyme. This antiserum was used successfully for the detection of the human PGMl polymorphism in fetal tissue from forensic casework samples, and the detection of low levels of PGM activity in rabbit haemolysates but was not suitable for screening human haemolysate PGM. The antiserum did not bind either PGM2 or PGM3 isozymes, but recognised the milk (PGM4) isozymes. This led to detailed studies of the biochemical and genetical features of the milk PGM isozymes and a novel method for screening the PGM2 isozyme polymorphism. The sheep anti-rabbit PGM was used to purify small amounts of human PGMl from skeletal muscle for use as an immunogen. An alternative method for producing the PGM protein, using cloned protein DNA sequence and a lambda gtll expression vector was also investigated

Breast Location for De Novo Extramedullary Myeloid Sarcoma

Background: Myeloid Sarcoma (MS) is a rare hematologic cancer, which can occur as a breast mass to be distinguished from other non-hematopoietic tumors. Case Presentation: This report describe the unusual clinical history of a young woman diagnosed with MS. Radiotherapy/surgery alone may be inadequate, while chemotherapy and hematopoietic stem cell transplantation demonstrated to improve the prognosis for the isolated extramedullary localization. Conclusion: Performing a needle biopsy in order to exclude the diagnosis of a primitive breast disease is irreplaceable

The prevalence of specific phobia by age in an Italian nationwide survey: How much does it affect the quality of life?

Introduction: The study aimed to see if a community survey conducted by clinical interviewers with semi-structured psychiatric interviews shows lifetime prevalence rates of Specific Phobia (SP) similar to those found by surveys carried out by lay interviewers and if the high level of impairment found in SP may be confirmed. Methods: This is a community survey on an Italian nationwide sample randomly selected from registers of municipalities. Tools: semi-structured ANTAS psychiatric interview derived from the SCID-DSM-IV, carried out by clinicians (psychologists or physicians); Short Form Health Survey (SF-12) as a measure of Quality of Life (QoL). Analyses: means of the χ2 test odds ratios were adopted to test several associations regarding SP prevalence. One-way ANOVA was used to compare different groups on attributable burden due to SP and/or other disorders in worsening QoL. Results: The lifetime prevalence of SP was 2.3%. No difference was found by age class. Females showed more than twice the frequency of males (p<0.0001). The disorders showing the closest association with SP were: social phobia (OR=17.53); general anxiety disorder (OR=11.57); anorexia (OR=11.13) and agoraphobia (OR=10.03), but also obsessive compulsive disorders (OR=8.8), eating disorders (OR=7.2), panic disorder (OR=5.9), post-traumatic stress disorder (OR=5.8), and major depressive disorder (OR=4.8) presented an association that achieved statistical significance. The QoL of people with SP and at least one disorder of anxiety, mood or eating in comorbidity, measured as a score at SF12, was worse than controls without SP (p <0.001) but that of people with SP without co-morbidity was not (p = 0.809). Conclusion: An epidemiological study conducted by clinical interviewers through semi-structured interviews appears to re-dimension the impact of SP, at least from the public health perspective. Future prospective studies will better clarify the role of SP in the context of anxiety disorders

The Burden of Comorbidity Between Bipolar Spectrum and Obsessive-Compulsive Disorder in an Italian Community Survey

© 2020 Carta, Fineberg, Moro, Preti, Romano, Balestrieri, Caraci, Dell’Osso, Disciascio, Drago, Hardoy, Roncone, Minerba, Faravelli and Angst. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY: https://creativecommons.org/licenses/by/4.0/). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.Background: The impact of the comorbidity between Obsessive-Compulsive Disorder (OCD) and Bipolar Disorder Spectrum (BDS) remains to be clarified. The objective of this study was to examine the lifetime prevalence of OCD, the strength of the association of OCD with comorbid BDS and the role of comorbidity of OCD with BDS in the impairment of health-related quality of life (HRQoL) in an Italian community survey. Methods: The study is a community survey. The sample (N = 2,267; women: 55.3%) was randomly selected after stratification by sex and four age groups from the municipal records of the adult population of one urban, one suburban, and at least one rural area in six Italian regions. Physicians using a semi-structured interview (Advanced Tools and Neuropsychiatric Assessment Schedule, ANTAS-SCID) made Diagnostic and Statistical Manual of Mental Disorders – 4th revision (DSM-IV) diagnoses of OCD, Major Depressive Disorder (MDD) and Bipolar Disorder (BD). HR-QoL was measured by the Health Survey Short Form (SF-12). Lifetime Hypomania and subthreshold hypomania were screened by the Mood Disorder Questionnaire (MDQ). BDS was defined as the sum of people shown to be positive for hypomania by the MDQ—with or without a mood disorder diagnosis—plus people with a BD-DSMIV diagnosis even if negative for hypomania at the MDQ. Results: Overall, 44 subjects were diagnosed with OCD, 6 with MDD and 1 with BD. The lifetime prevalence of OCD was 1.8% in men (n = 18) and 2.0% in women (n = 26). MDD with lifetime subthreshold hypomania (i.e., people screened positive at the MDQ, even without diagnosed mania or hypomania at the interview) was associated with OCD (OR = 18.15, CI 95% 2.45–103.67); MDD without subthreshold hypomania (and screened negative at the MDQ) was not (OR = 2.33, CI 95% 0.69–7.01). People with BDS were strongly associated with OCD (OR = 10.5, CI 95% 4.90–12.16,). People with OCD and BDS showed significantly poorer HR-QoL than people with OCD without BDS (F = 9.492; P < 0.003). Discussion: The study found a strong association between BDS and OCD. BDS comorbid with OCD was associated with more severe impairment of HR-QoL than OCD without comorbid BDS. Identification of symptoms of hypomania, including subthreshold symptoms, may therefore be important in people with OCD as they might predict a course with poorer HR-QoL.Peer reviewedFinal Published versio

The Use of Antidepressant Drugs and the Lifetime Prevalence of Major Depressive Disorders in Italy

BACKGROUND: The increased use of antidepressant drugs (ADs) improved the response to the needs of care although some community surveys have shown that subjects without lifetime psychiatric diagnosis (anxiety/depression) used ADs. OBJECTIVES: To evaluate the appropriateness and amount of prescription of psychotropic drugs in people with lifetime diagnosis of Major Depressive Disorder (MDD) by means of community survey with a semi-structured interview as a diagnostic instrument, administered by clinicians. METHODS: STUDY DESIGN: community survey. STUDY POPULATION: samples randomly drawn, after stratification from the adult population of municipal records. Sample size: 4.999 people were drawn in 7 centres of 6 Italian regions. TOOLS: questionnaire on psychotropic drug consumption, prescription, health services utilization; Structured Clinical Interview for DSM-IV modified (ANTAS); Training: interviewers were trained psychologists or medical doctors. RESULTS: 3.398 subjects were interviewed (68% of the recruited sample). The lifetime prevalence of DSM-IV MDD was 4.3% in males and 11.5% in females; antidepressant drugs were taken by 4.7% of subjects, 2.9% male and 5.9% female. 38% of males and 57% of females with lifetime diagnosis of MDD were taking ADs. CONCLUSIONS: Compared with studies using lay interviewers and structured tools the prevalence of the MDD was quite lower; ADs use was higher and tallied well with the data regarding antidepressant sales in Italy; the correspondence between lifetime diagnosis of MDD and ADs use was closer

De Novo Donor-Specific HLA Antibodies Developing Early or Late after Transplant Are Associated with the Same Risk of Graft Damage and Loss in Nonsensitized Kidney Recipients

De novo posttransplant donor-specific HLA-antibody (dnDSA) detection is now recognized as a tool to identify patients at risk for antibody-mediated rejection (AMR) and graft loss. It is still unclear whether the time interval from transplant to DSA occurrence influences graft damage. Utilizing sera collected longitudinally, we evaluated 114 consecutive primary pediatric kidney recipients grafted between 2002 and 2013 for dnDSA occurrence by Luminex platform. dnDSAs occurred in 39 patients at a median time of 24.6 months. In 15 patients, dnDSAs developed within 1 year (early-onset group), while the other 24 seroconverted after the first posttransplant year (late-onset group). The two groups were comparable when considering patient- and transplant-related factors, as well as DSA biological properties, including C1q and C3d complement-binding ability. Only recipient age at transplant significantly differed in the two cohorts, with younger patients showing earlier dnDSA development. Late AMR was diagnosed in 47% of the early group and in 58% of the late group. Graft loss occurred in 3/15 (20%) and 4/24 (17%) patients in early- and late-onset groups, respectively (p = ns). In our pediatric kidney recipients, dnDSAs predict AMR and graft loss irrespective of the time elapsed between transplantation and antibody occurrence

The lifetime prevalence and impact of generalized anxiety disorders in an epidemiologic Italian National Survey carried out by clinicians by means of semi-structured interviews

Background: Generalized anxiety disorder (GAD) is one of the most reported diagnoses in psychiatry, but there is some discrepancy between the cases identified in community studies and those identified in tertiary care. This study set out to evaluate whether the use of clinicians as interviewers may provide estimates in a community survey close to those observed in primary or specialized care. Methods: This is a community survey on a randomly selected sample of 2338 adult subjects. The Advanced Neuropsychiatric Tools and Assessment Schedule (ANTAS) was administered by clinicians, providing lifetime diagnosis based on the DSM-IV-TR. Health-related quality of life (HR-QoL) was measured with the Short-Form Health Survey (SF-12). Results: Overall, 55 (2.3%) subjects met the criteria for GAD, with greater prevalence in women (3.6%) than in men (0.9%): OR = 4.02; 95%CI: 1.96–8.26. Up to 40% of those with GAD had at least another diagnosis of mood, anxiety, or eating disorders. The mean score of SF-12 in people with GAD was 32.33 ± 6.8, with a higher attributable burden than in other conditions except for major depressive disorder. Conclusions: We found a relatively lower lifetime prevalence of GAD than in community surveys based on lay interviewers and a structured interview. The identified cases of GAD showed a strong impact on the quality of life regardless of co-morbidity and high risk in women, suggesting a profile similar to the one identified from studies in primary and specialized care