Whole-exome sequencing in obsessive-compulsive disorder identifies rare mutations in immunological and neurodevelopmental pathways.

Studies of rare genetic variation have identified molecular pathways conferring risk for developmental neuropsychiatric disorders. To date, no published whole-exome sequencing studies have been reported in obsessive-compulsive disorder (OCD). We sequenced all the genome coding regions in 20 sporadic OCD cases and their unaffected parents to identify rare de novo (DN) single-nucleotide variants (SNVs). The primary aim of this pilot study was to determine whether DN variation contributes to OCD risk. To this aim, we evaluated whether there is an elevated rate of DN mutations in OCD, which would justify this approach toward gene discovery in larger studies of the disorder. Furthermore, to explore functional molecular correlations among genes with nonsynonymous DN SNVs in OCD probands, a protein-protein interaction (PPI) network was generated based on databases of direct molecular interactions. We applied Degree-Aware Disease Gene Prioritization (DADA) to rank the PPI network genes based on their relatedness to a set of OCD candidate genes from two OCD genome-wide association studies (Stewart et al., 2013; Mattheisen et al., 2014). In addition, we performed a pathway analysis with genes from the PPI network. The rate of DN SNVs in OCD was 2.51 × 10(-8) per base per generation, significantly higher than a previous estimated rate in unaffected subjects using the same sequencing platform and analytic pipeline. Several genes harboring DN SNVs in OCD were highly interconnected in the PPI network and ranked high in the DADA analysis. Nearly all the DN SNVs in this study are in genes expressed in the human brain, and a pathway analysis revealed enrichment in immunological and central nervous system functioning and development. The results of this pilot study indicate that further investigation of DN variation in larger OCD cohorts is warranted to identify specific risk genes and to confirm our preliminary finding with regard to PPI network enrichment for particular biological pathways and functions.

Brazilian psychiatric brain bank: a new contribution tool to network studies.

There is an urgent need for expanding the number of brain banks serving psychiatric research. We describe here the Psychiatric Disorders arm of the Brain Bank of the Brazilian Aging Brain Study Group (Psy-BBBABSG), which is focused in bipolar disorder (BD) and obsessive compulsive disorder (OCD). Our protocol was designed to minimize limitations faced by previous initiatives, and to enable design-based neurostereological analyses. The Psy-BBBABSG first milestone is the collection of 10 brains each of BD and OCD patients, and matched controls. The brains are sourced from a population-based autopsy service. The clinical and psychiatric assessments were done by an expert team including psychiatrists, through an informant. One hemisphere was perfused-fixed to render an optimal fixation for conducting neurostereological studies. The other hemisphere was comprehensively dissected and frozen for molecular studies. In 20 months, we collected 36 brains. A final report was completed for 14 cases: 3 BDs, 4 major depressive disorders, 1 substance use disorder, 1 mood disorder NOS, 3 obsessive compulsive spectrum symptoms, 1 OCD and 1 schizophrenia. The majority were male (64%), and the average age at death was 67.2 ± 9.0 years. The average postmortem interval was 16 h. Three matched controls were collected. The pilot stage confirmed that the protocols are well fitted to reach our goals. Our unique autopsy source makes possible to collect a fairly number of high quality cases in a short time. Such a collection offers an additional to the international research community to advance the understanding on neuropsychiatric diseases.

The rational use of fine needle aspiration biopsy (FNAB) in diagnosing thyroid nodules.

AIM: Fine needle aspiration biopsy (FNAB) plays a crucial role in the diagnosis of thyroid nodules and enables the number of surgical operations to be reduced. Theoretically, FNAB should be carried out on all nodules, though currently only those displaying certain characteristics are biopsied. Indeed, to perform FNAB on all nodules may be regarded as an excess of zeal. Therefore, it seems advisable that the endocrinologist should be able to confirm on the spot the necessity and utility of FNAB. METHODS: We evaluated on a sample of 263 consecutive requests (209 female, 57 male; age 56.7+/-13.7 years) for FNAB in 2004: 1) the appropriateness of the investigation, 2) expected efficacy, 3) practical efficacy, 4) efficiency. FNAB was performed under echo-guidance in accordance with the standard technique. In 50%, 36%, 6%, 3%, 2% and 1% of cases, the echographic diagnosis was of MNG, UNG, pseudo-nodular lesion in ATD, lymph-node, neck cyst, suspected parathyroid lesion and tumefaction of the salivary glands, respectively. A pre-FNAB clinical risk score was assigned to each case on the basis of clinical and echographic data, with a maximum possible score of 11. The results of FNAB were subdivided into 5 categories according to the criteria of the BTA (Thy1-Thy5). After FNAB, a decisional category was assigned, ranging from ''observation'' to ''surgery''; this was subsequently (7-18 months) compared with the management strategy adopted by the attending physician. Information was gathered by means of telephone enquiry. RESULTS: 1) Appropriateness: on the basis of clinical and echographic findings, FNAB was not judged appropriate in 24% of cases because of either the lack of confirmation of a significant target (34%) or a low pre-FNAB risk score (range 0-2) (66%). The decisional category was ''observation'' in 87% of cases and ''further investigation'' in 13%. 2) Expected efficacy: FNAB was performed in 76% of cases. The biopsies (3%) performed on swollen lymph-nodes and extra-thyroid neck tumefactions, in which biochemical evaluation was positive, proved to be diagnostic but not classifiable according to the BTA. In 82% of the remaining cases, the result was Thy2 (observation) or Thy 4-5 (surgery). Thy3 results (surgery) were rare (1%). Thy1 results (16%) were yielded by the aspiration of colloid cysts (29%), solid lesions (10%) characterised by means of PTH-FNAB and Tg-FNAB, nodules (9%) no longer detectable on repetition of FNAB, nodules (16%) in which FNAB was already a repetition of a non-diagnostic investigation (2003), and nodules (9%) in which the presence of normal thyrocytes, ''hot'' scintigraphic image and prior decision of the surgeon advised against repeating FNAB. Of the patients with Thy1 results, 26% refused to repeat FNAB. In all, 95% of FNAB supported by biochemical evaluation yielded results that usefully contributed to patient management. The correlation between pre-FNAB clinical risk and cytological score according to the BTA proved significant (P<0.001). No difference in diameter was recorded between nodules with adequate cytology (23.3+/-0.9 mm) and those with inadequate cytology (25.2+/-1.6 mm). 3) Practical efficacy: 75% of patients were reached by telephone. In most cases, observation was the most frequent clinical choice, after echography and/or FNAB. The decisional category assigned after FNAB correlated significantly (P<0.001) with the approach adopted by the attending physician. d) Efficiency: following FNAB, 11 patients were assigned to surgery. DTC was detected in 100% of these cases (1 follicular carcinoma, 1 insular carcinoma, 9 papillary carcinoma). The success of FNAB (9/11) in detecting lesions that proved malignant on histological examination (11/11) was significant (P<0.05). Of the 2 Thy 3 cases, 1 was follicular carcinoma and 1 was follicular adenoma with adjacent papillary carcinoma. The incidence of thyroid carcinomas in the population studied was 5.5%. CONCLUSIONS: 1) Together with clinical-biochemical evaluation, echo-guided FNAB re-mains the first-line diagnostic test in the management of thyroid nodules; 2) a pre-FNAB clinical risk score is useful in limiting the number of probably inappropriate investigations; 3) efficacy, in terms of cytology results that are useful for patient management after FNAB (and after biochemical evaluation, when indicated) is high, enabling patients to be stratified in classes with different subsequent pathways; 4) in the vast majority of cases, FNAB influences subsequent clinical decisions; 5) false negatives cannot be excluded, while false positives are practically nil; 6) further indications may be yielded by studies on larger populations, and new prospects may emerge from the application of other techniques associated to FNAB.

Safety and efficacy of administering 0.2 mg of recombinant human TSH for two consecutive days as an adjuvant to therapy with low radioiodine doses in elderly out-patients with large nontoxic multinodular goiter.

AIM: The aim of this study was to evaluate the efficacy of recombinant human TSH (rhTSH) as an adjuvant to radioiodine therapy for nontoxic multinodular goiter (MNG) in elderly subjects. METHODS: Twelve elderly out-patients with large MNG (group 1) were studied. The effect of adjuvant rhTSH administration (0.2 mg i.m. on 2 consecutive days) before low-dose (131)I was compared with that of radioiodine alone in 8 out-patients matched for age and MNG volume (group 2). The follow-up period was similar in both groups. RESULTS: The number and severity of side-effects during the first month of treatment were similar in both groups. On final examination, the number of patients symptomatic for goiter was significantly lower in group 1 than in group 2 (P=0.03). In group 1, TSH levels peaked at 40.3+/-9.5 mU/L on day 3, from the baseline value of 0.5+/-0.1 mU/L (P<0.001). In group 2, baseline TSH was 0.4+/-0.1 mU/L. Although a marked increase in f-T3, f-T4 and Tg (P<0.001) was noted in both groups during the first 2 weeks of treatment, peak values were much higher in group 1 than in group 2. On final examination, a slightly significant increase (P=0.01) in TSH levels from the baseline was noted in both groups (group 1: 1.2+/-0.2 mU/L; group 2: 1.4+/-0.3 mU/L). The percentage of patients who did not need therapies to control TSH secretion at the last examination was higher in group 1 (83%) than in group 2 (38%). Only in group 1, a significant reduction was noted in mean anterior-posterior lobar width (31+/-1.7 mm) from the baseline value (24.5+/-1.7 mm, P=0.04). Thyroid volume was reduced from 78.1+/-11.7 mL to 49.4+/-13.4 mL (P=0.001) in group 1 and from 89.8+/-25.2 mL to 67.1+/-20.5 mL (P=0.04) in group 2. Six months after (131)I therapy, slight changes in thyroid length and tracheal lumen were noted in both groups. CONCLUSIONS: This long-term controlled study demonstrates that 0.2 mg of rhTSH on 2 consecutive days increases the efficacy of ambulatory (131)I dosages in treating nontoxic MNG in elderly subjects. Adequate drug preparation generally prevents side-effects due to short-term but marked thyrotoxicosis that is aggravated by rhTSH administration. An increase in thyroid volume reduction seems to be the most important effect of rhTSH administered before (131)I.

A case-controlled study on the quality of life in a cohort of patients with history of differentiated thyroid carcinoma.

Although quality of life (QoL) has become an important aspect of cancer rehabilitation, psychometric studies on thyroid cancer patients are rare. We performed a case-controlled study on QoL in patients with differentiated thyroid carcinoma (DTC). QoL was evaluated in 61 patients with a history of DTC diagnosed from < 1 to 23 yr earlier. An undetectable thyroglobulin (Tg) level after recombinant human TSH (rhTSH) testing was considered the best predictor of cure. QoL was evaluated by means of a general psychiatric interview, the self-rating Kellner Symptoms Questionnaire (KSQ) and the Hamilton Depression Scale (HDS). QoL was also evaluated in a control group of subjects on L-T4 therapy with a non-toxic multinodular goiter diagnosed from < 1 to 25 yr earlier. DTC and control subjects were similar in age, male-female distribution and concomitant psychiatric therapies. Per-week dosage of L-T4 was higher in DTC patients than in controls (p < 0.01). In neither group of subjects was there any correlation between current TSH levels or interval from diagnosis and KSQ or HDS scores. Only in DTC patients was there a positive correlation between age and KSQ (p < 0.05) or HDS (p < 0.01) scores. There was a significant difference in overall KSQ scores between DTC (33.4 +/- 2.1) and control (24.5 +/- 1.9; p < 0.01) subjects. The subscales of KSQ showed a significant inter-group difference. HDS scores were higher in DTC subjects (35.8 +/- 1.0) than in controls (30.0 +/- 1.1; p < 0.01). HDS score was significantly (p = 0.02) higher in female than in male DTC patients. In patients with papillary carcinoma there was a positive correlation between the MACIS (metastases, age, completeness, invasiveness, size) score and KSQ (p = 0.01) or HDS (p < 0.01) scores. After rhTSH testing, detectable Tg levels were found in 13% of DTC patients. In Tg-positive patients, KSQ and HDS scores were not different from those of Tg-negative patients. After an 8-14 month period, a significant decrease in the KSQ scale somatization (p = 0.02) was found in a sub-set of 31 DTC patients. In conclusion, even in the age of rhTSH testing, DTC patients suffer an impairment of their QoL, as noted when short-term L-T4 withdrawal was the gold standard. Longitudinal evaluation seems to indicate a slight improvement in QoL when safe rhTSH testing is extensively used in the management of the disease.

Effects of recombinant human thyrotropin on heart rate variability and blood pressure in patients on l-thyroxine-suppressive therapy for differentiated thyroid carcinoma.

BACKGROUND: Recombinant human thyrotropin (rhTSH) is now currently used for the follow-up of patients with differentiated thyroid carcinoma (DTC) after total thyroid ablation. Side effects after rhTSH could involve the autonomic system and TSH receptors are possibly expressed in the heart and coronary arteries. METHODS: Heart rate variability (HRV), studied by power spectral analysis of low (LF) and high frequency (HF) powers, blood pressure (BP) and their responses to orthostatism were investigated before and 3, 6, 9 days after the first of two administrations of rhTSH on alternate days in 11 patients on chronic l-thyroxine (l-T4) suppressive therapy for DTC and in 31 healthy controls. RESULTS: A transient asymptomatic decrease in systolic and mean BP was observed during the rhTSH test, but rhTSH did not modify sympathovagal control of HRV and the lying to standing responses. Decreased LF power and LF/(LF + HF) and LF/HF ratios in DTC patients versus healthy controls indicated a sympathetic failure ascribed to the TSH-suppressive therapy with l-T4 rather than to direct effects of rhTSH. CONCLUSIONS: These findings allowed us to confirm the cardiovascular safety of rhTSH and the absence of its effects on sympathovagal control of HRV when used in the follow-up of patients with normal heart function after thyroid ablation for DTC.

Bilateral carcinoma in situ of the testis and cystic fibrosis transmembrane conductance regulator (CFTR) mutation in an azoospermic patient with late-onset 21beta-hydroxylase deficiency.

Testicular cancer can impair spermatogenesis. In addition, chemotherapy or radiotherapy used for its treatment further damage testicular function mainly affecting highly proliferating germ cells. The multifaceted etiology of male infertility includes, among others, alterations of male reproductive tract differentiation such as monolateral or bilateral congenital absence of vas deferens and perturbations in adrenal steroid synthesis on a genetic basis such as 21beta-hydroxylase deficiency. Herein, we report the case of a male patient with primary infertility, probably related to a combination of genetic and acquired factors with different expressions over time.

A simple and safe nomogram for the management of oral anticoagulation prior to minor surgery.

In 80 consecutive, anticoagulated patients scheduled for minor surgery, we reduced warfarin daily dosage by 50% on days 4, 3 and 2 before the surgery, restoring the original dose the day immediately before surgery. The evening after surgery, patients took a double warfarin dose, and then the usual maintenance dose was reintroduced. The mean International Normalized Ratio (INR) value assessed 1 week before surgery was 2.63 (range 1.88-3.87); it decreased at the moment of performing surgery to 1.68 (range 1.42-2.20; P < 0.05 with respect to the preoperative value), and returned to 2.43 7 days after (range 1.96-3.51, P = ns with respect to the preoperative value). No significant difference was found comparing prothrombin fragment 1.2 (F1.2) levels 1 week before surgery and on the morning of surgery (0.49 ng/ml vs 0.67 ng/ml, P = ns), suggesting that no activation of blood coagulation had taken place following the reduction of anticoagulant therapy. Patients developed neither major nor minor bleeding, nor thromboembolism during the procedures or up to 1 month after surgery. In our experience, this method for the management of anticoagulation before minor surgery has been shown to be safe and useful, avoiding the cumbersome shift to either intravenous or subcutaneous heparin.