Validation Checklist for Massive Vaccination Centers.

Abstract: Starting from the minimum requirements indicated by Lombardy Region, a validation checklist has been developed by experts in design, healthcare layout planning, hygiene and public health, planning and compliance, in order to provide managers of COVID-19 massive vaccination centers with a useful and easy-to-use tool to ensure quality, safety and efficiency of the different activities performed.

Myelography in calves: a comparative study between the contrast agents iopamidol and iohexol / Mielografia em bezerros: estudo comparativo entre os meios de contrastes iopamidol e ioexol

Due to the scarcity of myelogenous studies in cattle, the present study aimed to evaluate the efficacy and distribution of iopamidol and iohexol contrast agents in calves, in order to determine guidelines for obtaining diagnostic radiographs of spinal cord disorders in these animals. Ten healthy Holstein calves, seven days to two months of age, were divided into two groups, according to the contrast medium applied. Myelographic studies of the spine were performed with the calves in lateral recumbency, with radiographs repeated 20 times during a two-hour period. On the radiographs, the contrast medium was analyzed for opacity, detail of the image, distension of the medullary canal, and progression of the contrast line. After seven days, the myelographic studies were repeated, with the contrast media exchanged between the groups. There were no significant differences in the quality of the images and speed of the spinal column filling between the two contrast media. Furthermore, the best quality radiographic images were obtained six to eight minutes after injection of the contrast in the cervical spinal segment, 80 minutes in the thoracic, and 20 minutes in the lumbar, sacral, and cauda equina segments.(AU)

Devido à escassez de estudos mielográficos em bovinos e relatos de complicações no procedimento, o presente estudo teve por objetivo avaliar a eficácia e a distribuição dos meios de contraste iopamidol e ioexol em bezerros, a fim de nortear a melhor conduta para o diagnóstico de afecções vertebrais e medulares nesses animais. Foram utilizados 10 bezerros Holandeses, hígidos, com idade entre sete dias e dois meses, distribuídos em dois grupos, conforme o meio de contraste aplicado. O estudo mielográfico da coluna vertebral foi realizado na posição laterolateral, repetido em 20 momentos, durante o período de duas horas. Nas radiografias, analisou-se o meio de contraste quanto à opacidade, detalhes da imagem, distensão do canal medular e progressão da linha de contraste. Após sete dias, foi realizado o segundo período experimental, que compreendeu a troca do meio de contraste dentro de cada grupo. Não houve diferenças significativas em relação à qualidade da imagem e à velocidade do preenchimento da coluna medular entre os dois meios de contraste. A partir da administração dos meios de contraste, a obtenção de imagens radiográficas de melhor qualidade deu-se após seis a oito minutos no segmento medular cervical, 80 minutos no torácico e 20 minutos nos segmentos lombar, sacral e cauda equina.(AU)

Clinical and radiographic changes of carpi, tarsi and interphalangeal joints of beef zebu bulls on semen collection regimen / Mudanças clínicas e radiográficas das articulações carpo, tarso e interfalangeana de touros de corte zebuínos em regime de colheita de sêmen

Osteoarthritis and osteochondrosis are highly correlated to reproductive failure in bulls. This study aimed to evaluate the carpal, tarsal and interphalangeal lesions in beef zebu bulls on semen collection regimen. Twenty-one beef cattle bulls, in a total of forty-one animals, were split into three age-based groups: animals from two to four years old (GI), from more than four to eight years old (GII) and above eight years old (GIII). The clinical findings were conformational changes of limbs, synovial effusion, peripheral venous engorgement of joints and prolonged decubitus. The total population showed moderate clinical manifestation and radiographic score. The GIII presented more severe joint lesions. Carpi and tarsi regions had discrete to difuse osteophytosis, subchondral cysts, cartilaginous flaps, bone incongruence and fragmentation, osteitis, and ankylosis. Interphalangeal joints presented osteophytosis, distal phalanx osteitis and enthesophytosis. The digital radiographic examination allowed full identification of articular lesions and their clinical correspondences, besides the positive correlation between age, body weight and radiographic score.(AU)

Osteoartrite e osteocondrose estão altamente correlacionados à falha reprodutiva em touros. Este estudo teve como objetivo avaliar as lesões do carpo, tarso e interfalangianas em touros zebuínos de corte em regime de coleta de sêmen. Vinte e um bovinos de corte, touros, provenientes de um total de 41 animais, foram divididos em três grupos baseados na idade, sendo eles: animais de dois a quatro anos (GI), de mais de quatro a oito anos (GII) e acima de oito anos de idade (GIII). Os sinais clínicos encontrados foram mudanças de conformação dos membros, derrame sinovial, ingurgitamento venoso periférico das articulações e decúbito prolongado. A população total avaliada apresentou manifestação clínica e escore radiográfico de grau moderado. Os touros mais velhos apresentaram maior gravidade nas lesões articulares. O GIII foi o que apresentou as lesões articulares mais severas. Identificou-se nos carpos osteofitose de discreta a difusa, cistos subcondrais, "flaps" cartilaginosos, fragmentação óssea, perda de congruência óssea, osteíte e anquilose; as lesões társicas obedeceram ao mesmo padrão. As articulações interfalangeanas apresentaram somente osteofitose, osteíte nas falanges distais e entesiofitose. O exame radiográfico digital permitiu a identificação completa das lesões articulares e suas correspondências clínicas, além da correlação positiva entre a idade, o peso corporal e o escore radiológico.(AU)

Acid-base balance in sheep with experimentally induced acute ruminal lactic acidosis / Equilíbrio ácido-base em ovinos com acidose láctica ruminal aguda induzida experimentalmente

This study aimed to investigate the changes in the acid-base balance of sheep with experimentally induced acute ruminal lactic acidosis (ARA). Ten ewes orally received 15 grams of sucrose per kilogram of body mass. Arterial blood samples for blood gas analysis were obtained at the following intervals: before the induction of ARA (control), and 2, 4, 6, 8, 10, 12, 16, 20, 24, 28, 32, 36, 48, 72, 96, 120 and 144 hours after sucrose administration. Urine samples for pH measurement were obtained at the following times: -15 days, -7 days, and immediately before sucrose administration, then at 24, 48, 72, 96, 120 and 144 hours. Thereafter, both blood and urine samples were obtained on the 2nd, 3rd, and 4th following weeks. From 4 hours after the induction, elevation of the pH, bicarbonate and base excess on the arterial blood was observed. After 12 hours, the animals showed a decrease of these parameters, as well as urine acidification, which are symptomatic of metabolic acidosis. Within 28 hours, all parameters were normalized except the base excess, which only returned to normal after 72 hours. Despite the occurrence of acidemia, there was no need for medication and no animals died.(AU)

Este trabalho objetivou estudar as alterações referentes ao equilíbrio ácido-base de ovinos com acidose láctica ruminal aguda (ARA) induzida experimentalmente. Dez ovelhas receberam oralmente 15 gramas de sacarose por quilograma de peso corporal. Amostras de sangue arterial para realização da hemogasometria foram obtidas nos seguintes intervalos: antes da indução da ARA (controle), duas, quatro, seis, oito, 10, 12, 16, 20, 24, 28, 32, 36, 48, 72, 96, 120, 144 horas após a administração da sacarose. Amostras de urina para análise do pH urinário foram obtidas nos seguintes momentos: 15 dias, sete dias e imediatamente antes da administração da sacarose, 24, 48, 72, 96, 120 e 144 horas. Posteriormente, tanto as amostras de sangue quanto de urina foram obtidas na segunda, terceira e quarta semanas seguintes. A partir de quatro horas da indução, elevação do pH, do bicarbonato e excesso de base no sangue arterial foram observados. Após 12 horas, os animais apresentaram diminuição dos parâmetros acima citados, como também acidificação da urina, que são sintomas de acidose metabólica. Decorridas 28 horas, todos os parâmetros retornaram à normalidade, exceto o EB, que somente retornou à normalidade após 72 horas. Apesar da ocorrência da acidemia, não houve necessidade de tratamento medicamentoso e nenhum animal veio a óbito.(AU)

Pattern of drug use among preterm neonates: results from an Italian neonatal intensive care unit.

BACKGROUND: Drug use in preterm neonates admitted to Neonatal Intensive Care Unit (NICU) has been investigated, so far, in terms of unauthorized or off-label use; very little is known on the use of combinations of different active substances, which is frequently required in this population (prophylaxis of infections, treatment of concomitant diseases). The aim of this study was to describe the most common patterns of drug use in an Italian NICU, focusing on those with nephrotoxic potential. METHODS: Medical records of preterm neonates (<37 weeks of gestational age) weighing less than 1,500 g at birth and admitted to an Italian NICU were scrutinized in a 3-year retrospective investigation. Analysis included drug exposure, duration of therapies, co-administration of drugs with potential renal side effects; also daily protein supplement was calculated from parenteral nutrition. RESULTS: A cohort of 159 preterm neonates was selected; 68 were born weighing less than 1,000 g (extremely low birth weight infants, ELBW, Group A), 91 weighed between 1,000 and 1,500 g at birth (Group B). Compared to Group B, neonates of Group A were more likely to receive pharmacological treatments: the most used drugs were antibiotics (especially ampicillin and amikacin, p = .07 and p < .001, respectively), antifungals (especially fluconazole, p < .001), and diuretics (especially furosemide, p < .001). Analysis of co-administration of drugs with potential nephrotoxicity showed ampicillin and amikacin as the most reported combination (94.1% of Group A and 31.9% of Group B), the combination of furosemide with antibacterials (ampicillin or amikacin) was also frequently reported, with average period of combination shorter than 2 days. CONCLUSIONS: ELBW infants were exposed to a higher number of drugs compared to other neonates and were more likely to receive associations of drugs with nephrotoxic potential (e.g. furosemide and amikacin), though only for short cycles. Further studies should evaluate the safety profile (especially potential renal side effects) related to most commonly used combinations.

Donepezil plasma concentrations, CYP2D6 and CYP3A4 phenotypes, and cognitive outcome in Alzheimer's disease.

PURPOSE: The purpose of the study is to evaluate whether donepezil (D) plasma concentrations and activity of CYP2D6 and CYP3A4 are associated with the therapeutic response of patients with mild to moderate Alzheimer's disease (AD). METHODS: This study comprised 54 patients affected by probable AD in therapy with D 10 mg/daily for at least 3 months. Plasma concentrations of D and its three main metabolites (6DD, 5DD, DNox) were assayed with a novel high performance liquid chromatography (HPLC) technique. Cognitive progression was assessed at baseline and at 9 months of follow-up with the mini mental state examination (MMSE). The activities of the two cytochromes involved in D metabolism-CYP2D6 and CYP3A4-were evaluated according to their metabolic ratios in plasma or urine, after test doses of probe drugs (dextromethorphan and omeprazole). RESULTS: A significant correlation was found between plasma levels of D and variations in MMSE scores after 9 months of therapy (r (2) = 0.14; p = 0.006). Neither the concentrations of D metabolites nor the metabolic ratios of CYP2D6 and CYP3A4 showed any correlations with cognitive variations. Low CYP2D6 activity and advanced age were associated with high D concentrations. Patients who were treated with CYP2D6 and P-glycoprotein (P-gp) inhibitors also had higher D plasma levels (mean difference = 19.6 ng/mL; p = 0.01) than those who were not. CONCLUSIONS: D plasma concentrations, but not cytochrome phenotyping, are associated with cognitive outcomes in AD patients.

QUANTITATIVE ANALYSIS OF PERIODONTAL PATHOGENS IN PERIODONTITIS AND GINGIVITIS.

Periodontal tissues surround the teeth and provide their attachment. Periodontal diseases include a mild and reversible form named gingivitis and periodontitis that is the main cause of tooth loss in adults. Gingivitis, that affects gums and coronal junctional epithelium, as well as periodontitis, that is characterized by loss of connective tissue attachment, are caused by a persistent inflammatory response promoted by alteration of periodontal biofilm. The aim of the study was to test whether the prevalence or relative amount of each species was associated with a particular clinical condition. Periodontal evaluation of 539 unrelated patients was performed by the Periodontal Screening and Recording (PSR) system. Subgingival samples were obtained from the site with the worst PSR score. A selection of eleven bacterial species was evaluated by quantitative real time PCR. Some bacterial species were found to be associated with all phases of periodontal disease, such as Tannerella forsythia, Treponema denticola, and Treponema lecithinolyticum, while other species were more specifically associated with periodontitis, such as Porphyromonas endodontalis and Porphyromonas gingivalis, or with gingivitis, such as Capnocytophaga ochracea and Campylobacter rectus. Quantitative and qualitative analyses helps to better understand the microbial changes associated with different stages of periodontal disease.

E-CADHERIN CODING GENE (CDH1) AND NONSYNDROMIC CLEFT LIP WITH OR WITHOUT CLEFT PALATE: IS THERE ANY ASSOCIATION?

Epithelial to Mesenchymal Transition (EMT) is an important process involved in cancer, embryogenesis and organ development. Its role in nonsyndromic cleft lip with or without cleft palate (NSCL/P) has been extensively investigated and successfully linked to the disease. In this study, we focused on a gene, CDH1, encoding for E-cadherin, a key protein in EMT. We carried out an association study on an Italian sample group, genotyping four single nucleotide variations within the CDH1 gene, in order to verify the potential role of this gene in NSCL/P etiology. Neither the haplotype nor the family-based association test revealed any association between the genotyped SNPs and the pathology. Our results demonstrate that, in our Italian sample study, the analyzed single nucleotide polymorphisms are not associated to NSCL/P.

INTERLEUKIN-6 GENE POLYMORPHISM MODULATES THE RISK OF PERIODONTAL DISEASES.

Gingivitis and periodontitis are the two main periodontal diseases. Both are characterized by inflammation of the tissues surrounding the teeth but while tissue damages observed in gingivitis are mild and reversible, destruction caused by periodontitis is deeper and irreversible. Periodontal diseases and levels of degeneration of tissues surrounding teeth depend on several interacting endogenous and exogenous factors. Polymorphisms of genes encoding molecules that modulate the immune response and tissue homeostasis are the main causes of individual susceptibility to periodontal diseases. The aim of this study was to investigate IL6, IL10 and VDR gene polymorphisms in a large number of subjects affected by either gingivitis or chronic periodontitis. The sample included 750 Italian patients. We found that the rs1800795 SNP located in the IL6 gene promoter was strongly associated with the occurrence of both gingivitis and periodontitis. Indeed, homozygous individuals with variant allele appeared less-susceptible to both gingivitis OR=0.47 (95% C.I. 0.27-0.82) and periodontitis OR=0.36 (95% C.I. 0.21-0.64). No evidence of association between periodontal diseases and IL10 or VDR polymorphisms was obtained. This data confirmed the role of IL6 in susceptibility to periodontitis among the Italian population. The evidence that IL6 polymorphisms are also involved in gingivitis has implications in periodontal disease pathogenesis and reduces the appeal of IL6 as a periodontitis biomarker.

Bio-Revitalization: Effects of NASHA on Genes Involving Tissue Remodeling.

BACKGROUND: The "non-animal stabilized hyaluronic acid" (NASHA) is a widely used product in bio-revitalization injective procedures in esthetic medicine. The present research aimed to quantitatively evaluate the therapeutic effect of one of the more used bio-revitalization products on cultured dermal fibroblasts. RT-PCR was used for gene expression profiling of some proteins known to be relevant in skin homeostasis. METHODS: Human dermal fibroblasts were seeded on a culture medium enriched with a product for dermal bio-revitalization, consisting of stabilized hyaluronic acid gel 20 mg/ml. After 24, 48, and 72 h of exposure, the cDNA was amplified by real-time PCR. Gene expression was quantified with the delta/delta calculation method. RESULTS: In this study, the gene of metalloproteinase (MMP)-13 is strongly expressed after NASHA incubation. The MMP-2 encoding gene instead is less expressed, but both evidence the same temporal trend, being progressively up-regulated after 24 and 48 h, thereafter the expression decreases, whereas MMP-3 maintains the same up-regulation at 72 h. Hyaluronan synthase 1 and desmoplakin are progressively up-regulated and increase at 24, 48, and 72 h. Hyaluronidase 1 and neutrophil elastase genes are overexpressed, but at 72 h they both exhibit the same behavior as the other degradative enzymes MMP-13 and MMP-2. CONCLUSIONS: Skin bio-revitalization by injecting the tested NASHA gel produces an enhancement in the expression of some genes involved in extracellular matrix degradation and organization. In this study, a time-dependent behavior, different for genes encoding degradative compared to synthesis proteins, was demonstrated.

Impact of pericardium bovine patch (Tutomesh(®)) on incisional hernia treatment in contaminated or potentially contaminated fields: retrospective comparative study.

PURPOSE: This retrospective comparative study analyzes the outcome of patients affected by incisional hernia in potentially contaminated or contaminated field, treated by three operative techniques. METHODS: 152 patients (62 M:90 F; mean age 65 ± 14 years) underwent incisional hernia repair (January 2002-January 2012) in complicated settings. Criteria of inclusion in the study were represented by the following causes of admission: mesh rejection/infection, obstruction without gangrene but with possible peritoneal bacterial translocation, obstruction with gangrene, enterocutaneous fistula or simultaneous presence of ileo- or colostomy. The patients were divided into three groups: A (n = 76), treated with primary closure technique; B and C (n = 38 each), with reinforcement by synthetic or pericardium bovine mesh (Tutomesh(®)), respectively. The prosthetic groups were divided into Onlay and Sublay subgroups. RESULTS: Significant decreases in C vs A were observed for wound infection (3 vs 37%) and recurrence (0 vs 14%), and in C vs B for wound infection (3 vs 53%), seroma (0 vs 34%) and recurrence (0 vs 16%). Patients with concomitant bowel resection (BR) (43%) showed (all P < 0.05) an increase of overall morbidity (55 vs 33%) and wound infection rate (42 vs 24%) compared to cases without BR. Morbidity presented no significant differences in C-Onlay or Sublay subgroups. B-Sublay subgroup has (all P < 0.05) lower overall morbidity (20 vs 75%), wound infection (10 vs 68%) and seroma (0 vs 46%) than B-Onlay. CONCLUSIONS: The pericardium bovine patch seems to be safe and effective to successfully repair ventral hernia in potentially contaminated operative fields, especially in association with bowel resection.

WITHDRAWN: Short- and long-term efficacy of cholinesterase inhibitors in older adults with alzheimer's disease and mixed dementia: results of a 21-month observational study.

Ahead of Print article withdrawn by publisher.

Changes in the activity and expression of protein phosphatase-1 accompany the differential regulation of NHE3 before and after the onset of hypertension in spontaneously hypertensive rats.

AIM: The Na(+) /H(+) exchanger NHE3 activity decreases in the proximal tubule of spontaneously hypertensive rats (SHRs) as blood pressure increases, and this reduction is correlated with higher NHE3 phosphorylation levels at the PKA consensus site serine 552. This study tested the hypothesis that this lowered NHE3 activity is associated with an increase in PKA activity and expression, and/or a decrease in protein phosphatase-1 (PP1) activity and expression. METHODS: Proximal tubule NHE3 activity was measured as the rate of bicarbonate reabsorption by stationary microperfusion. NHE3 phosphorylation and protein expression were determined by immunoblotting. PKA and PP1 activities were determined using specific substrates under optimal enzymatic conditions. RESULTS: The PKA activator, 6-MB-cAMP, increased the phosphorylation levels of NHE3 at serine 552 in the renal cortex; this increase happens to a much greater extent in young pre-hypertensive SHRs (Y-SHRs) compared to adult SHRs with established hypertension (A-SHRs). Likewise, the inhibitory effect of 6-MB-cAMP on NHE3 transport activity was much more pronounced in the proximal tubules of Y-SHRs than in those of A-SHRs. Renal cortical activity of PKA was not significantly different between Y-SHRs and A-SHRs. On the other hand, Y-SHRs exhibited higher protein phosphatase 1 (PP1) activity, and their expression of the PP1 catalytic subunit PP1α in the renal cortex was also higher than in A-SHRs. CONCLUSION: Collectively, these results support the idea that the lower NHE3 transport activity and higher phosphorylation occurring after the development of hypertension in SHRs are due, at least in part, to reduced PP1-mediated dephosphorylation of NHE3 at serine 552.

Psychometric properties of the pain assessment in advanced dementia scale compared to self assessment of pain in elderly patients.

AIM: The aims of the present study were to report on the psychometric properties of the Pain Assessment in Advanced Dementia (PAINAD) scale by comparing it with the gold standard method of self-reporting on a numerical rating scale (NRS), and to provide a categorical version of the PAINAD scale comparable with the verbal descriptor scale of the NRS. METHODS: Six hundred elderly patients with various degrees of cognitive impairment consecutively admitted to the acute geriatric section at Padua University were evaluated. Cognitive, functional, and health statuses were evaluated using the Mini-Mental State Examination (MMSE), activities of daily living, and the Cumulative Illness Rating Scale (CIRS), respectively. Pain measurements were obtained by administering the NRS and the PAINAD scale. RESULTS: Cognitive decline was recorded in 310 subjects (52%). The internal reliability of the PAINAD scale was adequate for all items, both in patients with dementia (α = 0.90) and in those with no cognitive impairment (α = 0.94). The psychometric evaluation demonstrated a stronger level of concurrent validity (Kendall's τ = 0.73, p < 0.0001) and inter-rater agreement (ĸ = 0.74, p < 0.0001) for the PAINAD compared with the NRS. CONCLUSION: Our findings clearly indicate that the PAINAD scale is a reliable and easily administered tool for assessing pain intensity also in elderly patients with advanced dementia.

Nutritional predictors of cognitive impairment severity in demented elderly patients: the key role of BMI.

INTRODUCTION: The body mass index (BMI) is commonly used to assess nutritional status and the Mini Mental State Examination (MMSE) is a validated tool for assessing cognitive status in elderly people. Nutritional and cognitive aspects are closely related in dementia. OBJECTIVES: To establish whether BMI predicts cognitive decline in demented patients and whether an "alarm" BMI cut-off exists for declining MMSE scores. SUBJECTS AND METHODS: 82 elderly demented patients underwent clinical, bio-chemical and functional assessment. DESIGN: Transversal study. RESULTS: The mean BMI was 26.08±4.48 kg/m² and the mean MMSE 18.68±5.38. Patients with BMI<25 kg/m² had significantly lower MMSE scores (16.5±5.53 vs 20.38±4.64; p 0.001), fat-free mass (FFM; 27.76±8.99 vs 37.38±10.58 kg; p<0.001), fat-free mass index (FFMI; 11.52±3.03 vs 14.67±2.89 kg/m²; p<0.001), and fat mass (FM; 24.90±6.89 vs 36.86±6.77 kg; p<0.001), as well as lower Mini Nutritional Assessment (MNA) scores (23.80±2.50 vs 25.00±2.29; p=0.03) and higher vitamin B12 levels (460.95±289.80 vs 332.43±82.07 pg/ml; p=0.01). In the sample as a whole, MMSE scores significantly correlated with scores for MNA (r=0.27, p=0.01), FFM (r=0.27, p=0.01), BMI (r=0.19, p=0.05), ADL (r=0.28, p=0.01) and instrumental activities of daily living (IADL; r=0.34, p=0.002). On multiple logistic regression, BMI<25 kg/m² was independently associated with the risk of moderate-severe cognitive impairment (OR=2.96; 95% CI; 1.16-7.55) and female gender was independently associated with severity of dementia (OR=3.14; 95% CI; 1.09-9.03). CONCLUSION: BMI seems to indicate global health status in elderly demented people and a BMI of 25 kg/m² can be considered an "alarm" cut-off, lower values coinciding with a worse cognitive status based on MMSE scores.

Evidence of LEF1 fetal-maternal interaction in cleft lip with or without cleft palate in a consistent Italian sample study.

Epithelial mesenchymal transformation is considered a cardinal process in orofacial development. Several molecular players appear to be involved in this delicate mechanism; the activation of LEF1 transcription factor by transforming growth factor beta 3 seems to be a key step for the correct flow of events. The failure of orofacial processes during embryonic development may provoke cleft lip and/or cleft palate malformations. The scope of the present investigation was to verify whether genetic variants at LEF1 could influence the risk of orofacial clefting. The approach was a family based association study involving a total of 512 Italian patients and their parents, 401 having cleft lip with or without cleft palate (CL/P) and 111 with cleft palate only (CPO). Haplotype association analysis provided moderate evidence of an association with clefting (p 0.01). A log-linear likelihood-based method was used to verify maternal and foetal-maternal association. An association between the maternal genotype and the occurrence of CL/P was observed at two polymorphic loci, at rs10022956 (P = 0.0049) and rs10025431 (P = 0.0065) respectively, while a foetal-maternal effect modulating the risk of clefting was found at locus rs10025431 (P = 0.0071). These data further corroborate the importance of the mother's genotype with regard to susceptibility to malformations and early-onset diseases.

Study of the 12q13 region in nonsyndromic cleft lip with or without cleft palate.

The 12q13 region has been suggested as a candidate locus for orofacial cleft by different investigators. In the present study we tested the region for linkage with non syndromic cleft lip with or without cleft palate in a collection of 39 Italian multigenerational families, using microsatellite markers. No evidence of linkage was detected between the marker map and NSCLP under different mode of inheritance nor with a nonparametric method. Formal level of linkage exclusion, were obtained for each point of the map. Genetic heterogeneity and the different impact of the candidate locus among populations could explain conflicting results obtained in different studies.

No association between polymorphisms in cubilin, a gene of the homocysteine metabolism and the risk of non-syndromic cleft lip with or without cleft palate.

Epidemiological studies have correlated lower maternal periconceptional levels of plasma folate and cobalamin with increased risk of delivering offspring presenting congenital malformations such as cleft lip with or without cleft palate (CL/P) or neural tube defects. A number of genetic studies aimed at correlating these biochemical levels or the occurrence of malformations with specific genetic defects or polymorphisms have been successfully performed. The cubilin gene (CUBN) codes for a carrier that plays a crucial role in cobalamin cell internalization. CUBN polymorphisms were previously found to be associated with spina bifida occurrence. In this work, a family-based association study was performed to test CUBN involvement in CL/P. A sample of 391 CL/P triads was investigated with three single nucleotide polymorphisms mapping on the cubilin gene. Association tests indicated no significant association between CL/P and marker alleles or marker haplotypes. No evidence of maternal effect and imprinting were obtained. These data suggest that CUBN is not involved in CL/P onset in the investigated Italian population.

Study of ABCB1 multidrug resistance protein in a common orofacial malformation.

The onset of embryonic malformations is greatly determined by the intrauterine environment, conditioned by maternal lifestyle, diet, drugs and medication intake, in addition to both foetal and maternal genotypes. Maternal C677T MTHFR genotype has been identified as important factor in cleft lip with or without cleft palate (CL/P) etiology. In the present study we evaluated the possible interaction between maternal methylenetetrahydrofolate reductase (MTHFR) and foetal ABCB1 genotypes. ABCB1 gene codes for a drug-transport pump in charge to protect the cell by extruding a variety of harmful exogens, but with a reduced activity in a folate-restricted condition. Maternal 677T genotype is translated in a reduced folate availability for the developing embryo who consequently may becomes more exposed to external insults. A family based association analysis was performed to test the effect of ABCB1 polymorphisms in clefting, in the whole sample and in the stratified sample accordingly to maternal MTHFR genotype. No evidence of association between ABCB1 polymorphisms and CL/P was detected. This suggests that ABCB1 or ABCB1-MTHFR feto-maternal interaction could have no effect in orofacial clefting or could play a role in a limited number of cases.

Evidence of an involvement of TFAP2A gene in non-syndromic cleft lip with or without cleft palate: an Italian study.

Unraveling of factors involved in multifactorial diseases is a great challenge. Different approaches can be contemplate and applied to a variety of congenital malformations. In the present investigation TFAP2A has been considered a good candidate gene for nonsyndromic cleft lip with or without cleft palate (NSCLP) aetiology, basing on a sum of considerations. TFAP2A has been seen involved in orofacial development in mice; it is located in the NSCLP candidate region 6p24; it codes for a transcription factor which regulates expression of IRF6, a gene implied in NSCLP; finally, it is embroiled in the branchiooculofacial syndrome, that includes clefting as feature. A family based association analysis was performed with a sample study of 405 NSCLP triads. Evidence of association was obtained with both single marker and haplotype analyses, thus providing a support for TFAP2A in NSCLP aetiology.