Population-based trends in prenatal screening and diagnosis for aneuploidy: a retrospective analysis of 38 years of state-wide data.

OBJECTIVE: To analyse population-based trends over the entire history of prenatal testing for aneuploidy. DESIGN: Retrospective analysis of state-wide data sets. SETTING: Australian state of Victoria with ~70 000 annual births. POPULATION: All pregnant women undergoing invasive prenatal testing at <25 weeks' gestation from 1976 to 2013. METHODS: Analysis of three state-wide data sets: (1) Prenatal diagnosis data set of 119 404 amniocenteses and chorionic villus samplings from 1976 to 2013; (2) central serum screening laboratory data set from 1996 to 2013; (3) government birth statistics from 1976 to 2013. MAIN OUTCOME MEASURES: Annual numbers and uptake rates of invasive prenatal tests and serum screening, indications for invasive prenatal testing, prenatal diagnoses of aneuploidy, diagnostic yield of invasive tests. RESULTS: Annual numbers of invasive prenatal tests climbed steadily from 1976, then declined from 2000. In 2013, the number of invasive prenatal tests was the lowest in 25 years, while the number of trisomy 21 diagnoses was the highest ever recorded. Annual uptake of serum screening climbed from 1.6 to 83% over 1996-2013. Results from 2013 showed a high diagnostic yield (15.8%) for a low rate of invasive testing (3.4% of births). Over four decades, the number of invasive procedures performed for each diagnosis of major chromosome abnormality declined from 100 to six. CONCLUSIONS: This study demonstrates historic reductions in the proportion of women undergoing invasive testing and dramatic improvements in diagnostic yield. Monitoring the impact of new prenatal technologies on this progress remains an important research priority. TWEETABLE ABSTRACT: Invasive prenatal testing has reached historic lows due to dramatic improvements in Down syndrome screening.

[Life-threatening injuries of the subcutaneous ventral abdominal veins in dairy cows]. / Lebensbedrohliche Verletzungen der Bauchwandvenen bei Milchkühen.

Injuries of the superficial veins on the ventral abdomen of dairy cows can result in life-threatening haemorrhage. Treatment of these injuries can be difficult because of the characteristics of the veins. Five cows aged 6.5 to 12 years were referred to the Animal Hospitale Zurich, because of intermittent haemorrhage from a ventral abdominal vein. Four of the cows were near term and one had calved two weeks before referral. Intermittent bleeding was seen from the highly convoluted and dilated subcutaneous tributaries to the cranial superficial epigastric vein (milk vein). The haematocrit of 2 cows was markedly lower than normal. Emergency slaughter of one of the cows was carried out because of advanced age. The other 4 cows were sedated and after application of local anaesthetic, the wounds were debrided and sutured. Two of the cows required a blood transfusion before being operated. There were no postoperative complications. This type of injury seems to occur predominantly in pluriparous dairy cows close to term, in which the tributaries to the milk vein have undergone varicose changes. Treatment should consist of surgical resection of the injured tissue and suturing of the venous wall, subcutaneous tissues and skin.

[Bursitis with severe tendon and muscle necrosis on the lateral stifle area in cattle]. / Bursitis mit schwerwiegenden Sehnen- und Muskelnekrosen an der Aussenseite der Kniegelenke beim Rind.

In 21 animals, chronic swelling on the lateral aspect of the stifle also known as «perigonitis¼, «stable-syndrome¼ or «bursitis bicipitalis femoris¼ were evaluated. Ultrasonography showed increased fluid in the distal subtendinous bursa of the biceps femoris muscle and structural changes in the tendons, muscles, subcutis and fasciae. Soft tissue swelling and an irregular contour of the lateral tibial condyle were typical signs on radiographs. Macroscopic changes were found at the insertion of the biceps femoris muscle, the distal subtendinous bursa of the biceps femoris muscle, the lateral collateral ligament of the stifle, the origin of muscles on the lateral femoral condyle and the lateral tibial condyle. They mainly consisted of tendon and muscle tissue necrosis with granulation tissue. Histology revealed areas of coagulation necrosis in tendons and ligaments, in which occasionally Onchocerca spp. were seen. The severity of lesions correlated well with the clinical signs, which were associated with a poor prognosis in advanced cases.

Clinical findings and treatment in 63 cows with haemorrhagic bowel syndrome.

The clinical, haematological and biochemical findings, treatment and outcome of 63 cows with haemorrhagic bowel syndrome are described. The general condition and demeanor were moderately to severely abnormal in all the cows. Signs of colic occurred in 27 cows, decreased rectal temperature in 46 and tachycardia in 44. With the exception of one cow, intestinal motility was decreased or absent. Transrectal palpation revealed dilatation of the rumen in 47 cows and dilatation of the small intestine in 18. Faecal output was markedly reduced or absent, and the faeces were dark brown to black and contained blood. Nine cows were euthanized immediately after physical examination. Conservative medical therapy was instituted in two cows; however, both were euthanized a few days later because of deterioration in condition. Exploratory right flank laparotomy was carried out in 52 cows. Of these, 22 were euthanized intraoperatively because of severe lesions. In 27 cows, intestinal massage to reduce the size of blood clots was carried out; 11 were euthanized several days postoperatively because of deterioration in condition. In three other cows, intestinal resection was carried out and all survived. Of the 63 cows, 19 (30.2%) survived and were healthy at the time of discharge from the clinic.

[Cystoscopy in a heifer with rupture of a patent urachus]. / Zystoskopie bei einem Rind mit Urachus persistens-Ruptur.

This case report describes the clinical, ultrasonographic and cystoscopic findings and treatment in a two-year-old Swiss Braunvieh heifer with rupture of a patent urachus. The lead signs in the seven-month-pregnant heifer were markedly abnormal general condition and demeanour and a pear-shaped abdomen. The heifer had severe azotaemia, and abdominal ultrasonography revealed ascites, which was diagnosed as uroperitoneum based on an elevated creatinine level in the fluid. A patent urachus was identified during cystoscopy; the endoscope could be advanced beyond the apex of the urinary bladder into the urachus. Based on all the findings, a diagnosis of uroperitoneum attributable to rupture of a patent urachus was made. The urachus was ligated twice via a left-flank laparotomy. The general condition normalised within a few days of surgery, and the patient calved normally and was in good health at follow-up evaluation.

[Haemorrhage caused by thrombocytopenia in 3 calves]. / Hämorrhagien bei 3 Kälbern infolge Thrombozytopenie.

The clinical findings and treatment of 3 calves with bleeding attributable to severe thrombocytopenia (1000 to 5000 platelets/microl) are described. The calves responded to treatment with whole blood transfusion, dexamethasone and other drugs. One calf was subsequently diagnosed with persistent bovine viral diarrhoea virus infection and was euthanased despite a favourable response to treatment. The other two calves were discharged after platelet counts had normalised and bleeding had stopped.

Radiographic Measurement of Hindlimb Digit Length in Standing Heifers.

The digital skeleton of the hindlimbs was evaluated radiographically in 27 standing Brown Swiss heifers. The lateral digital skeleton was significantly longer than its medial counterpart on both a hard (wooden block) and a softer (wooden block covered with a thin rubber mat) surface. There were no significant differences between lengths measured on the hard and the soft surface. The difference between the length of the lateral and medial digits originated at the level of the condyles and increased with P1 and P2. It was partially compensated by a higher third phalanx (P3) in the medial digit, but this did not offset the overall length difference. The findings of this study confirmed that the length asymmetry of the paired digits of cattle documented previously in post-mortem specimens is also present in living cattle. Further investigation is required to determine the clinical relevance of the length asymmetry of the digital skeleton in cattle.

[Pneumothorax during right-sided nephrectomy in a heifer]. / Pneumothorax als Komplikation einer rechtsseitigen Nephrektomie bei einem Jungrind.

A 250-kg heifer had signs of colic attributable to urolithiasis of the right kidney. Medical treatment did not result in resolution of clinical signs, and nephrectomy was carried out. The surgery was started with the heifer standing, and the 13th rib was resected. However, during blunt dissection of the kidney, air suddenly entered the pleural space and the heifer had acute severe dyspnoea. The hole in the pleural cavity was sutured and a chest drain was placed. Inhalation anaesthesia was then induced and nephrectomy could be completed without further complications. The heifer was discharged 11 days postoperatively, and was healthy and had been integrated into the herd 12 months after surgery. Pneumothorax must be considered a possible complication of rib resection in right-sided nephrectomy in cattle.

Differential disposition of lysophosphatidylcholine in diabetes compared with raised glucose: implications for prostaglandin production in the diabetic kidney glomerulus in vivo.

An early increased formation of renal prostaglandins in diabetes which follows the hydrolysis of cellular phospholipids by cytosolic phospholipase A2 is of considerable importance in determining subsequent cellular function. As the disposition of concomitantly formed lysophosphatidylcholine may also affect cellular function, we investigated the cellular fate of exogenous lysophosphatidylcholine in mesangial cell-enriched glomerular cores and showed that in cells taken from diabetic rats there is an increased net reformation of phosphatidylcholine. Positional distribution of labelled palmitate from sn-1 position palmitate-labelled lysophosphatidylcholine showed distribution to both sn-1 and sn-2 position of the phosphatidylcholine formed with a significantly increased sn-2 position labelling in diabetes. Although both a coenzyme A-dependent acyltransferase activity and a coenzyme A-independent transacylase activity could be shown in these cells, the increased phosphatidylcholine formation in cells taken from diabetic animals was due to an increase in coenzyme A-independent transacylase activity. By contrast, an increase in coenzyme-A independent transacylase activity could not be demonstrated in cultured mesangial cells maintained with prolonged raised glucose concentrations. Cell homogenates possess the ability to transfer fatty acid from lysophosphatidylcholine to lysophosphatidylcholine and lysophosphatidylethanolamine with subsequent formation of phosphatidylcholine and phosphatidylethanolamine, respectively. In preparations from diabetic animals phosphatidylethanolamine formed in this manner was increased in the presence of an inhibitor of cytosolic phospholipase A2, indicating that it may provide a substrate for phospholipase A2 activity; an effect not seen in cultured cells maintained at raised glucose concentrations. It is concluded that one effect of an altered disposition of lysophosphatidylcholine in cells from diabetic animals would be to spare fatty acids released following phospholipase A2 hydrolysis of phospholipid, possibly providing the substrate for prostaglandin production, an effect not seen with raised glucose alone.

[Asymmetry in digit length in cows with sole ulcer. A post-mortem study of slaughter cows]. / Längenasymmetrie der Zehen bei Kühen mit Rusterholz'schen Sohlengeschwüren. Eine Post-mortem-Untersuchung an Schlachtkühen.

OBJECTIVE: In this study, the disparity between the lengths of the lateral and medial digit in cows with a Rusterholz sole ulcer compared to non-affected cows was investigated. MATERIAL AND METHODS: In total, 100 hind feet were collected from slaughtered cows. Twenty-five feet had a pre-ulcerative lesion at the typical site, 26 had an ulcer < 1 cm² and 24 had an ulcer > 1.5 cm². Twenty-five normal feet were used as controls. The lengths of the metatarsal condyles and phalanges (Ph) were measured on plantarodorsal radiographs and three different overall lengths of the digits were generated. The software program SPSS was used for analysis. RESULTS: The mean lengths of Ph1 and Ph2 and the three mean overall lengths of the lateral digit were greater than those of the medial digit while the length of Ph3 was longer on the medial digit. With one exception, the comparison of overall length 2 of the pre-ulcerative lesion with the control group, the within-feet differences of the ulcer groups were not significantly different from the difference of the controls. CONCLUSION AND CLINICAL RELEVANCE: These results corroborate the length asymmetry of the bovine digits but further studies are needed to evaluate the effect of the asymmetry of the digits on the development of sole ulcers.

Arthrodesis of the proximal interphalangeal joints of a hindlimb in a heifer.

A two-year-old Braunvieh heifer was presented with a traumatic luxation of the second phalanx of the medial digit and concurrent subluxation of the second phalanx of the lateral digit of the right hindlimb. Closed reduction of both luxations was possible. Surgical arthrodesis was achieved using one narrow 4.5 mm three-hole equine locking compression plate for each joint. Placement of the bone plates resulted in stable arthrodesis of both proximal interphalangeal joints of the right hindlimb but there was persistent residual lameness. The heifer delivered a healthy calf but was slaughtered eight months after surgery because of varus deformity of the contralateral limb. Radiographs taken post-mortem revealed pronounced periosteal reactions involving both proximal interphalangeal joints and only partial bony bridging of the joint spaces.

Use of IgG- and IgM-specific ELISAs for the assessment of exposure status of chickens to Eimeria species.

Simple and reliable methods for the determination of the exposure status of chickens to Eimeria species are required. For this purpose an enzyme-linked immunosorbent assay (ELISA) detecting specific IgG and IgM antibodies in serum samples was evaluated. Sera from chickens hyperimmunized by intramuscular injection of a saline extract of Eimeria tenella sporozoites were used to determine optimal reaction conditions in the ELISA which were found to be at a serum dilution of 1:100 and an antigen concentration of 0.2 microgram per reaction well. Saline extracts of sporulated oocysts and purified sporozoites of E. tenella were also potent antigens but most studies were carried out with sporozoite antigen. In a trial with 80 chickens, concentrations of serum IgM directed against sporozoite antigen increased significantly 9 days after primary infection with 10,000 oocysts of E. tenella per animal. IgM levels subsequently decreased rapidly reaching a plateau level only slightly higher than uninfected controls by about 15 days post-infection. In chickens challenged with 10,000 oocysts 21 days after primary infection significant increases of IgM levels were observed 2, 6 and 12 days later. In contrast IgG levels increased only slightly after primary infection but significant increases occurred after challenge infection so that by Day 12 after challenge sporozoite-specific IgG levels were much higher than in control chickens. Thus, it may be possible to discriminate between chickens actually infected with Eimeria (as indicated by high levels of antiparasite IgM), chickens which have been repeatedly exposed to Eimeria (as indicated by high levels of antiparasite IgG) and unexposed birds. The applicability of this ELISA, using sporozoite antigen of E. tenella to practical situations was substantially confirmed, since sampling of over 1000 sera from commercially reared broilers and laying hens indicated that broilers, maintained on medicated food, had low levels of IgM and IgG whereas 84-97% of the laying hens, receiving drug-free feed, had relatively high IgG concentrations. These results reflect low and rare exposure to Eimeria infections in broilers and repeated exposure of the hens.

Length asymmetry of the bovine digits.

The lengths of the digital bones of the fore- and hind-limbs obtained post mortem from 40 cattle of different ages were measured using digital radiographs. The lengths of the individual digital bones and the overall length of the digit were determined using computer software. The lateral metacarpal/metatarsal condyle, and lateral P1 and P2 were significantly longer than their medial counterparts, whereas P3 of the medial digit was longer than its lateral partner. Measured from the cannon bone epiphysis to the tip of the pedal bone, the mean increased length of the lateral digit was 0.8 mm in the fore- and 1.5 mm in the hind-limb. When the lengths of the digital bones were summed, the mean length of the lateral digit was 1.8 mm longer in the fore-limb and 2.1 mm longer in the hind-limb. Based on these findings, it can be concluded that the lengths of the paired digits differ in cattle. The majority of cattle have longer lateral digits in the fore- and hind-limbs. This asymmetry might explain why the lateral hind-limb claws are predisposed to sole ulcers on hard surfaces. In the hind-limbs, the impact is transferred from the pelvis directly to the longer lateral digit. In the fore-limb claws, the tenomuscular attachment to the trunk may be involved in a more even weight distribution and in a shift of weight to the medial claw.

Small heat shock protein alteration provides a mechanism to reduce mesangial cell contractility in diabetes and oxidative stress.

BACKGROUND: Small heat shock proteins are expressed in many tissues and are proposed to regulate actin filament dynamics when dissociated into small aggregates and phosphorylated in a p38 mitogen-activated protein kinase (p38MAPK)-dependent manner. METHODS: p38MAPK activity and small heat shock protein-25 (Hsp25) were determined in glomeruli from rats with experimental diabetes induced by streptozotocin administration and in isolated glomeruli exposed to a free radical stress. Contractile responsiveness of mesangial cells was determined by the serum-induced contraction of cell-embedded type I collagen gels. RESULTS: In experimental diabetes, there is an activation of p38MAPK, a decrease in the size of Hsp25 molecular aggregates, from large to small homo-oligomers, and an increase in the phosphorylation of Hsp25. In control glomeruli, a free radical stress, H2O2, activated p38MAPK and increased Hsp25 in a concentration-dependent manner. Additionally, H2O2 decreased the contractility of cultured mesangial cells concomitant with an increase in Hsp25 phosphorylation and a reduction in Hsp25 aggregate size. These effects were significantly reduced by SB202190, an imidazole-derivative cell-permeable inhibitor of p38MAPK. CONCLUSIONS: It has been proposed that the generation of oxygen-derived free radicals in diabetes may be linked causally to a loss of glomerular contractile reactivity and thus hyperfiltration in the early stages of diabetes mellitus. This study provides a mechanism for alteration of mesangial cell contractile responsiveness through phosphorylation of Hsp25 and may be a mechanism underlying abnormalities in glomerular hemodynamics in diabetes and in the presence of free radical stress.

Association of cyclin-dependent kinase-4 and cyclin D1 in neonatal beta cells after mitogenic stimulation by lysophosphatidic acid.

Neonatal pancreatic islet beta cells retain a mitogenic capacity in response to growth factors. In this study an increased incorporation of 5-bromo-2'-deoxyuridine in response to oleoyl-lysophosphatidic acid is preceded by a GTP-dependent increase in phosphorylation of the extracellular signal-related kinase, ERK1. The presence of cyclin-dependent kinase-4 and an association with a catalytic partner cyclin D1, a process by which the progression through the cell cycle is regulated in other cell types, was shown to follow this. The mechanisms linking ERK1 phosphorylation and activation of cell cycle progression are not known. Investigation of this process in neonatal beta cells may provide a common pathway for the early response to growth factors and the conditions required for an increase in beta cell mass by proliferation.

Extracellular matrix components cooperate to activate phosphatidyl inositol-4-phosphate 5-kinase.

Interaction of mesangial cells with extracellular matrix proteins can provide a means to modify cellular anchorage and traction through an interaction of integrins with activation of the actin cytoskeleton. We investigated intracellular signalling of matrix components fibronectin and laminin in mesangial cells in monolayer and 3-dimensional configurations to show a fibronectin-dependent activation of phosphatidylinositol-4-phosphate 5-kinase (up to threefold), which is augmented by a laminin-dependent increase in phospholipase D activity. Functional responsiveness to fibronectin and laminin addition was seen in the contraction of free-floating 3-dimensional mesangial cell-embedded collagen gels, a well-defined system reflecting coupling of extracellular matrix-cell events to activation of the actin cytoskeleton. Activation of phosphatidylinositol-4-phosphate 5-kinase and contraction of mesangial cell-embedded collagen gels in response to fibronectin and laminin were inhibited by pretreatment of mesangial cells with lovastatin and restored by isoprenoid augmentation with geranylgeraniol, supporting a role for the ras-related protein Rho in this process.

Increased phosphorylation of focal adhesion kinase in diabetic rat kidney glomeruli.

Altered extracellular matrix production by the glomerular mesangium is a feature of diabetes mellitus. Matrix proteins, including fibronectin, via interaction with cell-surface receptors (the integrins) may activate intracellular pathways such as prostaglandin production, shown previously to be stimulated by addition of fibronectin to glomerular cores. However, the signalling pathways involved are unclear. An intracellular tyrosine kinase (focal adhesion kinase), associated with focal adhesions, is known to be phosphorylated after interaction with matrix proteins. We now show for the first time, in glomeruli from diabetic rats, that focal adhesion kinase has increased phosphorylation on tyrosine, when compared with non-diabetic control rats. This phosphorylation was labile and disappeared with extended time of sample preparation or digestion of glomeruli to glomerular cores. Cultured mesangial cells, from non-diabetic rats, plated onto fibronectin also showed increased tyrosine phosphorylation of focal adhesion kinase accompanied by a twofold increase in prostaglandin production. However, it may not be possible to replicate fully the diabetic ¿state¿ in vitro merely by use of raised glucose concentrations, as these conditions (for 3 weeks) resulted in decreased focal adhesion kinase phosphorylation, despite increased fibronectin and prostaglandin levels. A role for increased focal adhesion kinase phosphorylation in kidney glomeruli isolated from diabetic rats, and any linkage to intracellular signalling pathways remains to be determined.

Production of hyaluronan by glomerular mesangial cells in response to fibronectin and platelet-derived growth factor.

The formation of the non-sulphated glycosaminoglycan hyaluronan by cells of the renal glomerulus in diabetes may contribute to altered matrix composition. We describe an increased production of hyaluronan from mesangial cell-enriched glomerular cores from diabetic animals, and further show that increased hyaluronan production follows the exposure of non-diabetic and diabetic preparations to fibronectin and to platelet-derived growth factor in vitro. Hyaluronan production appeared dependent on protein kinase C activity, and could not be shown after prolonged phorbol ester preincubation. Stimulation by fibronectin was wholly dependent on cyclooxygenase activity and prior prostaglandin production, while the effect of platelet-derived growth factor showed only a partial dependence.