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The experiment described in this research communication aimed to compare the immunological traits of Simmental (sire) × Holstein (dam) crossbred cows with the two parental breeds in the peripartum and early lactation period and to estimate the effects of heterosis for these traits. Flow cytometric evaluation of leukocyte subpopulations was assessed in 16 Crossbred (CR), 8 Holstein (HO) and 8 Simmental (SI) cows. Estimated average values of innate and adaptive immune cells showed statistically significant differences between the crossbred cows and parental breeds. Interestingly, the most relevant differences between the three groups related to adaptive immune cells. In particular, the CR cows showed a lower percentage of CD3+ T lymphocytes compared with the SI group (P < 0.0001) and the highest proportions of CD21+ B lymphocytes among the three groups (P < 0.0001). Furthermore, we found the highest positive value of heterosis for the CD21+ B lymphocytes (7.0) and the lowest negative value for CD3+ T lymphocytes (−4.8) in F1 derived population. It seems reasonable to believe that these differences could affect immune function of crossbred cows.

The aim of the research reported in this paper was to evaluate plasma concentrations of energy, oxidative and inflammatory biomarkers of Simmental (sire) × Holstein (dam) crossbred cows, in comparison with the two parental breeds during the peripartal and early lactation periods and to estimate the effects of heterosis for these traits. Thirty-three animals, managed under the same conditions, 8 Simmental (SI), 9 Holstein (HO) and 16 crossbred (CR) cows were enrolled in this study. Glucose, non-esterified fatty acids (NEFA), β-hydroxybutyrate (BHB), total bilirubin, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), creatine kinase (CK), total protein, albumin, creatinine and urea were determined in blood sampled at six different time points (30 ± 3 and 15 ± 3 d before the expected calving date, at calving and 15, 30 and 60 d after calving). Furthermore, derived reactive oxygen metabolites (d-ROMs), biological antioxidant potential (BAP), interleukin-6 (IL-6), haptoglobin (Hp) and serum amyloid A protein (SAA) were determined to evaluate inflammatory and oxidative status. Results showed that the CR group had significantly lower average values of glucose and NEFA when compared to HO group; signifcantly lower values of urea than SI group and significantly higher values of creatinine than HO. Furthermore, CR cows showed the lowest average value of d-ROMs with respect to SI and HO parental breeds. Finally, the average value of haptoglobin was significantly lower in CR and HO groups, when compared to SI group. As for the heterosis we found the highest (positive) percentage for CK (98%) and BAP (47%) and the lowest (negative) percentage for OSi (−75%) and d-ROMs (−39%). A negative percentage was also found for the glucose (−11%) and NEFA (−20%) toward the Simmental parental breed. Our results suggest a different response among the three genetic groups during the peripartal and early lactation periods. In particular, CR and SI cows seem more adaptable regarding energy metabolism and oxidative status. Heterosis led to a positive effect on those parameters in Simmental (sire) × Holstein (dam) crossbred cows F1 population (50% Simmental and 50% Holstein).

Monocytes are bone marrow derived innate myeloid cells that circulate in the blood and play important roles in infection and inflammation. As part of the mononuclear phagocytic system, monocytes provide innate effector functions, support the adaptive immune response, and play a role in the maintenance of tissue homeostasis. In addition to their role in sensing pathogen-associated molecular patterns using several pattern recognition receptors, monocytes are characterized by their ability to ingest and kill microbes, to produce cytokines and chemokines, and to present antigens to T cells. For a long time, monocytes have been considered as a homogenous cell population, characterized by the expression of CD14, the receptor of lipopolysaccharide. Studies in several species have shown that the monocyte population consists of phenotypically and functionally different cell subsets. In this review, we report a comprehensive phenotyping of monocyte subsets in cattle. In addition, the most characterizing cell markers and gating strategies for detailed immunophenotyping of bovine monocyte subsets are discussed.

We present the case of a 51 years-old woman who was referred to our Liver Unit for suspected dysmetabolic liver disease. She had a previous diagnosis of Systemic Sclerosis and Sjögren’s Syndrome and had altered hepatic enzymes with a positive anti-Nuclear Antibodies centromeric pattern, anti-Cardiolipin Antibodies and anti b2 Glycoprotein I Antibodies. Despite complete liver assessment and disease staging was negative, the clinical course was complicated by the development of anemia, due to esophageal varices bleeding associated with worsening splenomegaly. Liver biopsy was key for reaching the diagnosis as it showed portal tracts with fibrous expansion, ductular proliferation and focal lympho-granulocytic infiltrate, reduced caliber of portal vessels, hypoplastic portal tracts, focal herniation, aberrant microvasculature and positive endothelial CD34 staining. Having ruled out any other cause of portal hypertension such as cirrhosis, blood diseases, occlusion of the hepatic and portal veins, etc., we finally concluded that the portal hypertension was due to hepatoportal sclerosis associated with Systemic Sclerosis and Sjögren’s Syndrome. Keywords: Idiopathic Non-Cirrhotic Portal Hypertension (INCPH; Hepatoportal Sclerosis (HPS); Systemic sclerosis (SSc); Sjögren’s Syndrome (SS).

Bovine intramammary infections are common diseases affecting dairy cattle worldwide and represent a major focus of veterinary research due to financial losses and food safety concerns. The identification of new biomarkers of intramammary infection, useful for monitoring the health of dairy cows and wellness verification, represents a key advancement having potential beneficial effects on public health. In vitro experiments using bovine peripheral blood mononuclear cells (PBMC), stimulated with the bacterial endotoxin lipopolysaccharide (LPS) enabled a flow cytometric assay in order to evaluate in vivo poly-ADP-ribose (PAR) levels. Results showed a significant increase of PAR after 1 h of treatment, which is consistent with the involvement of PARP activity in the inflammatory response. This study investigated PARP-1 activation in leukocyte subpopulations from bovine milk samples during udder infection. A flow cytometric assay was, therefore, performed to evaluate the PAR content in milk leukocyte subsets of cows with and without intramammary infection (IMI). Results showed that milk lymphocytes and macrophages isolated from cows with IMI had a significant increase of PAR content compared to uninfected samples. These results suggest mastitis as a new model for the study of the role of PARP in zoonotic inflammatory diseases, opening a new perspective to the “One Health” approach.

The main objective of this study was to assess PAG2 mRNA expression in maternal blood cells at the peri-implantation period in water buffalo, moreover, we wanted to evaluate the earliest time in which PAG-2 could be detected in maternal blood. Thirty-two lactating buffaloes artificially inseminated (AI) were utilized. Blood was collected at Days 0, 14, 18, 28, 40 after AI (AI = day 0). Pregnancy was diagnosed by ultrasound at Days 28 and 40 post AI. Out of 32 buffaloes, 14 were pregnant (P group) and 18 were not pregnant (NP group). The plasma PAG-2 threshold of 1.0 ng/mL in the P group was reached at day 40 post AI. PAG2 mRNA expression differed between the P and NP groups, and was either evaluated in Peripheral Blood Mononuclear Cells (PBMC) or Polymorphonuclear Leukocytes (PMN), starting from day 14. However, both the estimated marginal means and multiple comparisons showed that PAG2 mRNA expression was higher in PMN than PBMC. In the present study, PAG-2 appeared in the blood (40 Days post AI), and an early expression of PAG2 mRNA at Day 14 post AI was also observed. Although further research is undoubtedly required, PAG2 mRNA in peripheral blood leukocytes could be using to better understand the role that PAGs play during pregnancy in buffalo.

The identification of cross-reactive monoclonal antibodies (mAbs) that recognize orthologous leukocyte differentiation molecules (LDM) in buffaloes has overcome a major impediment limiting research on the immune response to pathogens and development of vaccines. As reported, two pilot trials were conducted to accomplish two objectives: (1) demonstrate that multiparameter flow cytometry can be conducted equally well in buffalo with mAbs directly and indirectly labeled with fluorochromes in research and (2) flow cytometry can be used to compare and extend studies on diseases of economic importance to buffalo using bovine viral diarrhea virus (BVDV) as a model pathogen. Pregnant buffalo cows were infected with BVDV-1 at 81 (trial 1) and 203 (trial 2) days post artificial insemination and flow cytometric evaluations were performed at 0, 3, 4, and 14 days after infection (dpi). Fluorochrome conjugated mAbs were used in trial 1, and fluorochrome conjugated goat isotype specific anti-mouse antibodies were used to label mAbs in trial 2. Flow cytometric analysis revealed a transient lymphopenia occurs during the 1st days following infection similar to lymphopenia reported in cattle. In particular, significant differences were observed between pre- and post-infection absolute values of T lymphocytes (−56%, P < 0.01). CD21+ B lymphocytes (−65%, P = 0.04), and Natural Killer cells (−72%, P < 0.001). No significant differences were observed in monocytes and neutrophil absolute values, or the CD4:CD8 ratio. Animal health status was followed until 15 days after calving. No clinical signs of infection were observed during the evaluation period, however, animals in trial 1 developed complications later the infection. One cow aborted at 57 days post-infection, the second cow developed a prolapse a day after calving and died. These two animals also showed a more pronounced lymphopenia in comparison with animals infected at 203 days of pregnancy (e.g., −77 vs. −22% T lymphocytes at 3 dpi, respectively). The pilot studies have demonstrated that it is possible to use multicolour multiparameter flow cytometry to study the immune response to pathogens affecting the health of buffalo.

Flow cytometry is a powerful technology used in many fields of cell biology. It is also used as a routine method to count somatic cells in milk and to characterize bovine milk leukocytes. In this study, we used flow cytometry to simultaneously assess the viability, the percentage of the single subsets of leukocytes and to quantify the expression of CD11b, an immunological marker of cell activation status. Immunological markers were then related with on farm recorded parameters as milk electrical conductivity (MEC) and average milk flow rate (MFR). Composite milk samples were collected from 43 cows, nine of which had naturally infected udders and 34 of which had no infected udders. First, the milk samples were classified according to bacteriological test in positive and negative. The results showed that the negative samples to bacteriological test had: (i) significantly higher percentages of live lymphocytes, (ii) significantly lower percentages of CD11b+ leukocytes, (iii) significantly lower MEC and higher MFR values. Then, samples were classified in three groups according to somatic cell count (SCC): Group A (n = 15), samples with SCC &le, 100,000 cells/mL, all negative to bacteriological analysis, Group B (n = 11), samples with 100,000 &lt, SCC &lt, 300,000 cells/m, with four samples positive to bacteriological analysis, Group C (n = 17), samples with SCC &ge, 300,000 cell/mL with five samples positive to bacteriological analysis. Multivariate discriminant analysis was used to verify which flow cytometry immunological markers and on farm recorded parameters could better discriminate among the different groups of SCCs. Linear discriminant analysis (LDA) indicated that 5 of the 10 parameters could best be used to reveal the differences between positive and negative samples among the considered groups of SCCs. Furthermore, the Canonical discriminant analysis (CDA) indicated that composite milk samples with different SCC and infection status were clearly separated from each other in a two-dimensional space. In conclusion, the study highlighted that: (1) the conventional flow cytometry analysis applied on milk samples is a useful tool to investigate immunological parameters as potential indicators of udder health, (2) the combined evaluation of live milk leukocytes and recorded farm parameters could be useful to assess udder health status in dairy cows. The results obtained from this pilot study on few data require new and larger trials to be confirmed.

Cluster of differentiation 4 (CD4) is the accessory protein non-covalently bounded to T cell receptor (TCR) that recognize an invariant region of MHC class II on antigen presenting cell (APC). Its cytoplasmic tail, physically associated with a protein tyrosine kinase, is important in the activation of helper/inducer T (TH) lymphocytes. The anti-bovine CD4 monoclonal antibody (mAb) CC8 is a clone usefully used in flow cytometry assay in the last 20 years and no anomalous labelling has been described in previous reports. The aim of this study was to investigate the lack and reduced binding of clone CC8 to TH lymphocytes in three different breeds of dairy cattle: Italian Friesian (FRI), Pezzata Rossa Friulana (PRI) and their crossbreed (MET). Two unexpected flow cytometric phenotypes were found: CC8+ (reduced labelling) in 25%, 35% and 15% of the animals in the three breeds respectively, and CC8− (complete lack of labelling) in 15% of PRI cows. The complete sequencing of CD4 coding region (CDS) has allowed to identify two new missense mutations in the exon 5: Q306H and K310 N associated with the two phenotypes. A further new synonymous mutation (g.103634725 G > A) was found in exon 6. Further studies are needed to understand the effects of the news mutations, but these results demonstrate that flow cytometry is a powerful tool to discover polymorphic determinants of antigens involved in the immune responses.

BACKGROUND Nasal congestion represents a troublesome health issue which is especially and invalidating in children. Effective nasal drugs, such as sympathomimetic drugs, are usually forbidden in children under 12 years of age because of their potential systemic adverse effects. Hypertonic nasal physiological solutions have recently been successfully used to decongest nasal mucosa in children: its mechanical activity has been universally recognized as safe and effective and it represents a well-established, useful treatment in children. METHODS We have retrospectively analyzed a case series of 40 children treated for 4 days (96 hours) with a new class 1s medical device nasal hypertonic spray containing Pirometaxine™ (Narlisim™) in outpatient affected by nasal congestion due to common cold. Every child was evaluated on a 3-point symptom assessment scale (0: no symptom; 1: mild symptom; 2: moderate symptom; 3: severe symptom) at the beginning of the trial (T0) and after 48 (T1) and 96 hours (T2). The symptoms assessed were nasal obstruction, nasal secretion, headache, flash of cold, pharyngodynia, cough, and sneeze. RESULTS The results, in terms of short-term efficacy to control nasal obstruction (T1 vs. T0: P

Experimental evidences support a direct role for leptin in immunity. Besides controlling food intake and energy expenditure, leptin was reported to be involved in the regulation of the immune system in ruminants. The aim of this work was to highlight the expression of leptin receptor (LEPR) on Bubalus bubalis immune cells using a multi-approach assessment: flow cytometry, confocal microscopy and gene expression analysis. Flow cytometric analysis of LEPR expression showed that peripheral blood monocytes were the predominant cells expressing LEPR. This result was corroborated by confocal microscopy and RT-PCR analysis. Moreover, among lymphocytes, LEPR was mainly expressed by B lymphocytes and Natural Killer cells. Evidence of LEPR expression on buffalo blood leukocytes showed to be a good indicator of the responsivity of these cells to leptin, so confirming the involvement of leptin in buffalo immune response.

Poly(ADP-ribosyl)ation (PAR) is a post-translational protein modification catalysed by enzyme member of the poly(ADP-ribose) polymerases (PARPs) family. The activation of several PARPs is triggered by DNA strand breakage and the main PARP enzyme involved in this process is PARP1. Besides its involvement in DNA repair, PARP1 is involved in several cellular processes including transcription, epigenetics, chromatin re-modelling as well as in the maintenance of genomic stability. Moreover, several studies in human and animal models showed PARP1 activation in various inflammatory disorders. The aims of the study were (1) to characterize PARP1 expression in bovine peripheral blood mononuclear cells (PBMC) and (2) to evaluate PAR levels as a potential inflammatory marker in cells isolated from blood and milk samples following different types of infection, including mastitis. Our results show that (i) bovine PBMC express PARP1; (ii) lymphocytes exhibit higher expression of PARP1 than monocytes; (iii) PARP1 and PAR levels were higher in circulating PBMCs of infected cows; (iv) PAR levels were higher in cells isolated from milk with higher Somatic Cell Counts (SCC > 100,000 cells/mL) than in cells from milk with low SCCs. In conclusion, these findings suggest that PARP1 is activated during mastitis, which may prove to be a useful biomarker of mastitis.

Leptin has a pleiotropic effect on regulating appetite, energy metabolism, growth, reproduction, body composition and immunity. This property supports leptin and its receptor as candidate genes for evaluating genetic polymorphisms to associate with growth, milk yield and other economic traits. The aim of this study is to characterize the leptin receptor gene in Bubalus bubalis, to identify single-nucleotide polymorphism (SNP) sites in different coding and non-coding regions and to analyse potential associations between SNPs identified and the body measurements traits of growing buffalo heifers. A group of 64 animals were genotyped by direct sequencing and twenty-eight SNPs were detected. A sequence analysis revealed the presence of nine interesting SNPs in gene sequence. The association analysis of polymorphisms with the body measurements traits of growing buffalo heifers shows significant statistical effects on chest depth and sacrum height. Therefore according to the results obtained from this study, the leptin receptor gene appears to have potential effects on the body measurement traits of Bubalus bubalis.

In this study, the effect of polymorphisms in the leptin gene on the hematological variables in periparturient dairy cows was investigated. The hematological profile of 67 Holstein cows was assessed for 6 wk around calving. The DNA of the cows was genotyped at 6 polymorphic loci within the leptin gene, and 7 haplotypes were reconstructed. Significant haplotype substitution effects were found, for haplotype 1, on total white blood cell count for 2 wk around calving (+0.70 10(3)/μL, P = 0.05; +1.38 10(3)/μL, P = 0.0001); on neutrophil cell count in the first week after calving (+0.94 10(3)/μL, P = 0.001); on lymphocyte count during the 3 wk before and the first week after calving (+0.32 10(3)/μL, P = 0.05; +0.27 10(3)/μL, P = 0.03; +0.26 10(3)/μL, P = 0.04; +0.34 10(3)/μL, P = 0.01); on red blood cell count during the last week before calving and wk 1 and 2 after calving (+0.21 10(6)/μL, P = 0.02; +0.23 10(6)/μL, P = 0.01; +0.20 10(6)/μL, P = 0.03); on mean corpuscular volume (-1.35 fL, P = 0.01; -1.29 fL, P = 0.002; -1.18 fL, P = 0.004; -1.09 fL, P = 0.008; -1.23 fL, P = 0.003; -1.31 fL, P = 0.003); and on mean corpuscular hemoglobin (-0.37 pg, P = 0.05; -0.38 pg, P = 0.02; -0.39 pg, P = 0.01; -0.34 pg, P = 0.03; -0.40 pg, P = 0.01; -0.40 pg, P = 0.01) during all 6 wk analyzed. Significant haplotype substitution effects, but opposite those of haplotype-1, were found for haplotype-2 on white blood cell count (-1.10 10(3)/μL, P = 0.01; -1.30 10(3)/μL, P = 0.002; -1.09 10(3)/μL, P = 0.01) and neutrophil count (-0.82 10(3)/μL, P = 0.02; -0.95 10(3)/μL, P = 0.005; -0.92 10(3)/μL, P = 0.01). Haplotype-3 influenced red blood cell count (-0.23 10(6)/μL, P = 0.03; -0.28 10(6)/μL, P = 0.01; -0.34 10(6)/μL, P = 0.002) during the last 2 wk before and the first week after calving, and also, with effects evident only in wk 3 and 2 before calving, mean corpuscular volume (+1.38 fL, P = 0.03; +0.97 fL, P = 0.05; +1.08 fL, P = 0.05), mean corpuscular hemoglobin (+0.58 pg, P = 0.02; +0.38 pg, P = 0.04; 0.51 pg, P = 0.01), and red blood cell distribution width (-0.56% P = 0.02; -0.47%, P = 0.05). The current study provided evidence that several polymorphisms in the leptin gene play a role in the variability of hematological variables during the peripartum period, and might be used as genetic markers for improving the immunological conditions of dairy cows in critical productive periods.

To date, only a few SNP in the leptin receptor gene have been detected and analyzed while the leptin gene has been extensively characterized and several polymorphisms were detected and associated to functional and productive traits in cattle. In this work, for the first time, the bovine leptin receptor gene was fully characterized by sequencing the whole coding region and part of the 5’ flanking region. A group of 95 Holstein cows was genotyped in order to search for SNP at both the LEPR and LEP genes, and to associate them with milk and morphological traits. Nine novel SNP in the leptin receptor gene and one novel SNP in the leptin gene were detected. Four SNP of the leptin receptor gene and one SNP of leptin gene showed a significant effect on one or more analyzed traits, and, in all cases, the greatest effect was observed on fat content. This study provided knowledge of the existence of further polymorphisms in the leptin receptor and leptin genes that have an influence on some important economic traits.

Insulin-like growth factor I (IGF-I) plays an important role in mammary growth and function. We detected two novel SNPs in the 5′UTR of the IGF-I gene: g.855G>C and g.857G>A of locus X69472, and genotyped 103 sheep of three breeds at the identified polymorphisms. The same animals were genotyped at a previously reported SNP (g.271C>T; accession X69473) located in exon 3. From the SNPs we built 4 haplotypes. We calculated lactation persistency from test day records of the genotyped animals as the ratio of milk (kg), fat (g) and protein (g) yield from day 86 to 170 of machine milking to yields during the first 85 days of machine milking. We estimated allele substitution effect on persistency for each SNP and haplotype. We found that allele T of SNP g.271C>T and haplotype [G;T] had a positive effect on maintaining a constant yield level during lactation. The improvement of lactation persistency could be highly beneficial to the sustainable use of local breeds.

One hundred seven cattles of four breeds (Holstein–Friesian, Jersey, Valdostana and Piedmontese) were genotyped at the DGAT1 K232A and the SCD1 A293V variants, as well as at 7 SNPs of the GHR gene, that were used to construct 14 GHR haplotypes. Allele frequency at most SNPs, and GHR haplotype frequencies highly differed between breeds. The effect of each SNP and haplotype was associated to milk production traits. The consistent effect of the 232K allele of DGAT1 on the increase of fat and protein content was confirmed in the Holstein and the Jersey, while this allele is missing in the Valdostana and rare (0.03) in the Piedmontese. SCD1 A293V variant showed no effect on either milk yield or fat and protein content. The positive effect of GHR F279Y variant on milk fat and protein content, although much weaker than the effect of DGAT1, was confirmed. The combination of the GHR SNPs into haplotypes seems to improve the association, likely for the interaction of variant F279Y with a SNP in the GHR gene promoter.

Summary The objective of this work was to characterize the complete coding region of the ovine acyl CoA:diacylglycerol acyltransferase 1 (DGAT1) gene of three Italian sheep breeds: Sarda, Altamurana and Gentile di Puglia. Characterization was accomplished by direct sequencing of 8676 bp of the relevant DNA, including introns and partial 5¢ and 3¢ untranslated regions (UTRs). We detected five novel SNPs; one SNP (g.5553C>T) is located in intron 2, has similar frequencies in the three breeds and showed a negative association with milk fat content. More interesting is an SNP in the 5¢ UTR (g.127C>A), the occurrence of which is rare in the higher milk-fat breeds (Altamurana and Gentile di Puglia); it is located in the core sequence of Sp1, a putative binding site of transcription factors. This SNP showed a significant negative association with milk fat content in the Sarda sheep. Because DGAT1 plays a fundamental role in triacylglycerol synthesis, the novel detected SNP in the 5¢ UTR of the DGAT1 gene might explain, at least partially, the variation of fat content in the milk of Sarda sheep.

Leptin is a protein involved in the regulation of feed intake, fat metabolism, whole body energy balance, reproduction and hematopoiesis. In cattle Leptin gene has been considered a potential QTL influencing several production traits like meat production, milk performance and reproduction. Several studies on bovine leptin gene have found association between polymorphisms and traits like milk yield, feed intake, fat content, carcass and meat quality. With the aim to assess the presence of sequences polymorphisms in the Buffalo leptin gene, we sequenced the entire coding region and part of the introns on a panel of Italian River Buffalos. In this study we identified a new set of SNP (Single Nucleotide Polymorphism) useful for association studies.

In this work, the genetic variation of milk FA was investigated in three different bovine breeds, the Jersey, the Piedmontese and the Valdostana, and at different lactation stages. All animals were genotyped for 21 Single Nucleotide Polymorphisms located within nine candidate genes involved in lipid synthesis: diacylglycerol acyltransferase 1 and 2 (DGAT1, 2); stearoyl-CoA desaturase (SCD); growth hormone receptor (GHR); fatty acid synthase (FASN); acyl-CoA dehydrogenase (ACAD); fatty acid binding protein (FABP4); lipoprotein lipase (LPL); and leptin gene (LEP). The highest milk-fat Jersey breed also showed the highest content of saturated FA. Throughout lactation, the breeds showed a similar variation in the FA, with a decrease in the short-chain, this was accompanied by a general increase in the long chain FA at the end of lactation. The increase in long chain saturated FA was particularly evident in the case of the Jersey. The effect ofSCDgene on the C14 desaturation index was confirmed; theDGAT1gene was polymorphic only in the Jersey breed, but its effect was confirmed only on milk fat content; three further potential candidate genes were identified: first, theFABP4gene, which was found to influence medium and long chain FA in all the breeds, but not the desaturation indices; second, theFASNgene, which was found to influence the amount of PUFA in the Piedmontese and the Valdostana, and third, theLPLgene, which was found to affect fat content in the Piedmontese.

This observational study was undertaken in maintenance renal transplant recipients to assess the relationship between cyclosporine C(2) and the long-term risk of chronic renal allograft dysfunction (CRAD). Pharmacokinetic profiling was undertaken twice yearly in 79 patients with stable graft function receiving cyclosporine microemulsion (Neoral) and steroids. Mean time since transplantation at study entry was 56 +/- 49 months posttransplant. At the end of the observational period (mean 43 +/- 14 months) 24 patients (30%) had developed CRAD, defined as proteinuria > 500 mg/24 hours associated with a rising serum creatinine and confirmed by graft biopsy. There were no significant differences at baseline between patients who did vs did not develop CRAD, except for reduced incidence of acute rejection in CRAD-free patients. The cyclosporine AUC was significantly higher among patients without CRAD (5707 ng. h/mL vs 3994 ng. h/mL, P =.0007). Mean C(2) was also significantly higher: 1001 ng/mL in the CRAD-free group versus 640 ng/mL in the CRAD group (P =.002). There was no significant difference in C(0). Regression analysis showed that the best predictors for the occurrence of CRAD were a low AUC (relative risk [RR] 1.54, P 900 ng/mL. C(2) appears to be a superior strategy to C(0) monitoring of cyclosporine in stable renal transplant patients with regard to the risk of CRAD. C(2) values above 900 ng/mL are appropriate to minimize the risk of CRAD among patients receiving cyclosporine microemulsion and steroids

The aim of this study was to evaluate the safety of conversion to sirolimus (SRL) immunosuppression among 19 renal transplant recipients (KTX) with progressive chronic renal allograft dysfunction (CRAD). Conversion to SRL was performed with concomitant sharp withdrawal of the calcineurin inhibitor (CI). SRL was added at a starting dose of 3 mg, then adjusted to obtain SRL target trough blood levels of 8 to 10 ng/mL. CI were stopped the evening before starting SRL. All patients enrolled in the study have now completed 6 months of follow-up: all are alive without acute rejection or major infection following rapid conversion to SRL. No significant change in the 6 months postconversion hematologic and hepatic profile was observed compared with the preconversion values, while significant dyslipidemia was induced. After conversion to SRL, significant amelioration of the renal function was found in 36% of patients, stabilization in 21%, and continuous deterioration in 43%. Patients whose renal function improved were found to have been converted at a significantly lower creatinine: (pre 2.6 +/- 0.9 vs post 1.9 +/- 0.2; P =.038) with respect to those patients who had continuous renal deterioration. In KTX with CRAD, sharp withdrawal of CI with concomitant conversion to SRL is safe, avoiding major infections, acute rejections, and significant side effects. Short-term amelioration of the renal function is best obtained when early conversion is performed. Long-term follow-up will be necessary to confirm these preliminary data.

A more precise determination of the date of primary rubella infection in pregnancy is obtained by application of the urea denaturation method. Two denaturation buffers containing 6 and 8 M urea were compared for the detection of rubella IgG avidity by enzyme immunoassay. The results demonstrate that IgG avidity detected by 6M urea increases more rapidly, from 16% on the third day to 51% on the 46th day after the rash, than that obtained by 8 M urea, from 13 to 36%. The specific rubella IgG avidity detected by 6 M urea denaturation buffer increased significantly in paired sera obtained at an interval of 15 days. The same samples did not show any significant increase of IgG avidity when an 8 M urea denaturation buffer was used. The use of a mild denaturant such as 6 M urea indicates the presumptive date of primary rubella infection in pregnancy within 2 months of sampling.

Tacrolimus, a macrolide derived from the fungus Streptomyces tsukubaensis, has been shown in clinical trials to effectively reduce the incidence of acute rejection in kidney transplantation. Analysis of data shows that the reduced incidence of moderate to severe acute rejection episodes with tacrolimus is irrespective of whether the diagnosis was based on clinical signs and symptoms, local biopsy reading, or blinded central review of the biopsies. The potent immunosuppressive action of tacrolimus prompted us with the idea to wean off corticosteroid therapy, a major cause of long- term complications in renal transplant recipients. The aim of this study was to evaluate safety and efficacy of withdrawing corticosteroid therapy 6 months after transplantation, in a group of 60 renal allograft recipients immunosuppressed with tacrolimus plus mycophenolate mofetil and steroid.

The aim of this study was to evaluate the efficacy of lithium prophylaxis during the peripartum period in women with lithium-responsive bipolar I disorder. Seventeen lithium-treated patients were selected and underwent precon- ception counseling that included both a psychiatric and an obstetric evaluation. Treatment was continued with flexible-doses of lithium combined with supportive psycho- therapy throughout the pregnancy and the postpartum period. The results support the prophylaxis efficacy of lithium in lithium-responder bipolar women who have a high risk of severe peripartum recurrences.

Pregnancy has been associated with a low risk of HIV disease progression. Most pregnancies with HIV currently involve women who have not experienced AIDS-defining events, and are clinically classified as Centers for Disease Control and Prevention (CDC) groups A or B. We evaluated the main maternal outcomes among pregnant women with more advanced HIV disease, defined by CDC-C disease stage. Data from the Italian National Program on Surveillance on Antiretroviral Treatment in Pregnancy were used. A total of 566 HIV-infected mothers, 515 in stage A or B (CDC-AB group) and 51 in stage C (CDC-C group) were evaluated. The two groups had similar baseline characteristics. No differences were found in the main maternal and neonatal outcomes. Most of the women achieved viral suppression at end of pregnancy (>1000 copies per milliliter: CDC-C: 17.2%; CDC-AB: 13.7%). One year after delivery, HIV replication (HIV-RNA >1000 copies per milliliter) was present in 11.5% of CDC-AB women and 30.0% CDC-C women. Despite lower initial CD4 counts (300 versus 481 cells per microliter), CDC-C women maintained stable CD4 levels during pregnancy, and 1 year after delivery, a significant increase in CD4 count from preconception values was observed in both groups (CDC-C: +72 cells per microliter, p=0.031; CDC-AB: +43 cells per microliter, p