Artificial Oxygen Carrier With Pharmacologic Actions of Adenosine‐5′‐Triphosphate,Adenosine, and Reduced Glutathione Formulated to Treat an Array of Medical Conditions

Effective artificial oxygen carriers may offer a solution to tackling current transfusion medicine challenges such as blood shortages, red blood cell storage lesions, and transmission of emerging pathogens. These products, could provide additional therapeutic benefits besides oxygen delivery for an array of medical conditions. To meet these needs, we developed a hemoglobin (Hb)‐based oxygen carrier, HemoTech, which utilizes the concept of pharmacologic cross‐linking. It consists of purified bovine Hb cross‐linked intramolecularly with open ring adenosine‐5′‐triphosphate (ATP) and intermolecularly with open ring adenosine, and conjugated with reduced glutathione (GSH). In this composition, ATP prevents Hb dimerization, and adenosine promotes formation of Hb polymers as well as counteracts the vasoconstrictive and pro‐inflammatory properties of Hb via stimulation of adenosine receptors. ATP also serves as a regulator of vascular tone through activation of purinergic receptors. GSH blocks Hb's extravasation and glomerular filtration by lowering the isoelectric point, as well as shields heme from nitric oxide and reactive oxygen species. HemoTech and its manufacturing technology have been broadly tested, including viral and prion clearance validation studies and various nonclinical pharmacology, toxicology, genotoxicity, and efficacy tests. The clinical proof‐of‐concept was carried out in sickle cell anemia subjects. The preclinical and clinical studies indicate that HemoTech works as a physiologic oxygen carrier and has efficacy in treating: (i) acute blood loss anemia by providing a temporary oxygen bridge while stimulating an endogenous erythropoietic response; (ii) sickle cell disease by counteracting vaso‐occlusive/inflammatory episodes and anemia; and (iii) ischemic vascular diseases particularly thrombotic and restenotic events. The pharmacologic cross‐linking of Hb with ATP, adenosine, and GSH showed usefulness in designing an artificial oxygen carrier for multiple therapeutic indications.     

Liposome‐Encapsulated Hemoglobin Improves Tumor Oxygenation as Detected by Near‐Infrared Spectroscopy in Colon Carcinoma in Mice

Liposome‐encapsulated hemoglobin (LEH) with high (h‐LEH, P₅₀O₂ = 10 mm Hg) or low O₂ affinity (l‐LEH, P₅₀O₂ = 40 mm Hg) may improve O₂ delivery to sensitize tumor tissues for radiotherapy. A total of 10 mL/kg of h‐LEH, l‐LEH, red blood cells (RBCs), or saline was infused in mice transplanted with murine colon carcinoma with near‐infrared spectroscopy (NIRS) detectors set at the tumor (right leg) and intact muscle (left leg). NIRS recorded changes in the amount of oxyhemoglobin (oxyHb), deoxyhemoglobin (deoxyHb), and their sum (tHb) with the animals spontaneously breathing room air (10 min), pure O₂ (5 min), and then back to room air. The tumor was finally excised for histological examination. In mice treated with h‐LEH, tHb significantly increased compared to mice receiving other solutions. The magnitude was significantly attenuated in the tumor compared to the intact muscle under room air. Reciprocal changes in oxyHb and deoxyHb between intact muscle and tumor in response to infused solutions allowed assumption of average tissue PO₂ between 30 and 40 mm Hg in muscle and at around 10 mm Hg in tumor. While O₂ respiration increased oxyHb and decreased deoxyHb both in muscle and tumor, their sum or tHb consistently decreased in muscle and increased in tumor regardless of preceding infusion. Such responses were totally reversed when mice were placed under hypoxia (10% O₂), suggesting that a lack of physiological circulatory regulation in tumor may account for heavier immunohistochemical staining for human hemoglobin in tumors of mice treated with h‐LEH than with l‐LEH. The results suggest that h‐LEH may cause significant tumor oxygenation compared to RBC, l‐LEH, or saline probably due to its nanometer size (vs. RBC) and high O₂ affinity (vs. l‐LEH) without increasing O₂ content in the intact tissue (vs. O₂ respiration) probably due to a lack of physiological circulatory regulation.     

Vancomycin hemodialysis: Clearance differences between high‐flux hemodialysis and on‐line hemodiafiltration

The aim of this study was to analyze the differences between vancomycin clearance (Kd) with high‐flux hemodialysis (HFHD) and on‐line hemodiafiltration (OL‐HDF). The OL‐HDF therapy combined the diffusion and convective transport of solutes. To compare the Kd, a vancomycin loading dose of 1 g was administered intravenously post‐dialysis to 11 chronic and anuric (<100 mL/24 h) hemodialysis patients, undergoing HFHD and post‐dilutional OL‐HDF in consecutive therapies. Additional doses of 0.5 g were administered after 45 minutes at the end of each dialysis therapy during antibiotic treatment. Blood samples were drawn from arterial and venous lines at the start of hemodialysis sessions and at the first, second, third, and fourth hours. Additional samples were drawn at 15, 30, and 45 minutes after the end of dialysis therapy. Vancomycin plasma concentration, blood urea nitrogen (BUN), creatinine, and β₂‐microglobulin were measured. The patients’ hydration status was evaluated by bioimpedance analysis. The mean of vancomycin dialyzer clearance (Kd_dc) calculated was 110.8 ± 15 mL/min with HFHD and 146.8 ± 13.8 mL/min with OL‐HDF (P = 0.025). Significant differences were also obtained for β₂‐microglobulin clearance, Kd_dc 72.6 ± 15.4 mL/min with HFHD and 113.4 ± 24.2 mL/min with OL‐HDF (P = 0.012), whereas no differences were found for BUN or creatinine. Additionally, to analyze differences between HFHD and OL‐HDF, a variable volume dual pool mathematical model was developed to estimate the body clearance (Kd_bc), extraction mass (M_e), and inter‐compartment mass‐transfer coefficient (K₁₂) of each molecule. A higher vancomycin Kd_dc with OL‐HDF produced by convection improved removal of antibiotic; this can compromise achieving a therapeutic concentration target. We recommended evaluating increased loading doses of vancomycin and avoiding administration during OL‐HDF to assure adequate treatment.     

Gasoline exhaust damages spermatogenesis through downregulating α6‐integrin and β1‐integrin in the rat model

The health influence of air pollution has been an international public health concern. Increasing evidence has suggested that air pollution has been associated with decreased sperm quality. However, the underlying molecular mechanisms are still not fully elucidated. We aimed to verify whether gasoline exhaust leads to reproductive impairment by injuring spermatogonial stem cells and explore its underlying molecular mechanism. Twenty male Sprague‐Dawley rats were randomly divided into two groups: the exposure group (n = 10) and the control group (n = 10). After 6‐month exposure, the sperm count and morphology were determined. The histological changes in the seminiferous tubules were examined by HE staining. The expression of α6‐integrin and β1 ‐ integrin was assessed with Quantitative RT‐PCR, Western blot and Immunohistochemical staining. Compared with control group, male rats exposed to gasoline exhaust showed significantly reduced sperm count, increased sperm abnormality rate and the total number of spermatogonia, primary spermatocytes, secondary spermatocytes, spermatids were decreased. (all p < .01). The expression levels of α6‐integrin and β1 ‐ integrin in the exposure group were significantly lower than those in the control group (all p < .01). Our study showed that exposure to gasoline exhaust caused impairment to spermatogonial stem cells through downregulating α6‐integrin and β1 ‐ integrin.     

CAG‐repeat polymorphisms in the polymerase γ gene and male infertility: a meta‐analysis

CAG‐repeat in the polymerase γ (POLG) gene encoding polymerase γ for mitochondria is important to spermatogenesis. Compared with a few researchers who raised alteration of CAG‐repeat‐affected male reproductive ability, others did not find the association between CAG‐repeat polymorphisms and male infertility. Therefore, a comprehensive meta‐analysis is necessary to determine the association; 13 case–control studies were screened out using keywords search. From these studies, characteristics were extracted for conducting meta‐analysis. Odds ratio (OR) and 95% confidence interval (CI) were used to describe the results; the results indicated that CAG‐repeat allele was not a risk factor to male infertility (pooled OR = 1.03, 95% CI: 0.79–1.34, P = 0.828). Four different genetic comparisons also demonstrated a negative result: heterozygote comparison (not 10/10 versus 10/10. Pooled OR = 0.99, 95% CI: 0.77–1.27, P = 0.948), homozygote comparison (not 10/not 10 versus 10/10. Pooled OR = 1.08, 95% CI: 0.56–2.06, P = 0.816), the recessive genetic comparison (not 10/not 10 versus not 10/10 + 10/10. Pooled OR = 1.07, 95% CI: 0.58–1.95, P = 0.829) and the dominant genetic comparison (not 10/not 10 + not 10/10 versus 10/10. Pooled OR = 0.97, 95% CI: 0.72–1.29, P = 0.804); based on current researches, this meta‐analysis demonstrated no apparent association between POLG‐CAG‐repeat and male infertility. Similarly, CAG‐repeat was not a sensitive site to male infertility.     

Liposome‐Encapsulated Hemoglobin Accelerates Skin Wound Healing in Diabetic dB/dB Mice

Since liposome‐encapsulated hemoglobin with high O₂ affinity (h‐LEH, P₅₀O₂ = 10 mm Hg) has been reported to accelerate skin wound healing in normal mice, it was tested in dB/dB mice with retarded wound healing, as seen in human diabetics. Two full‐thickness dorsal wounds 6 mm in diameter encompassed by silicone stents were created in dB/dB mice. Two days later (day 2), the animals were randomly assigned to receive intravenous h‐LEH (2 mL/kg, n = 7) or saline (2 mL/kg, n = 7). The same treatment was repeated 4 days after wounding (day 4), and the size of the skin lesions was analyzed by photography, surface perfusion was detected by Laser‐Doppler imager, and plasma cytokines and chemokines were determined on days 0, 2, 4, and 7, when all animals were euthanized for morphological studies. The size of the ulcer compared to the skin defect or silicone stent became significantly reduced on days 4 and 7 in mice treated with h‐LEH (47 ± 8% of original size), similar to the level in wild‐type mice, compared to saline‐treated dB/dB mice (68 ± 18%, P < 0.01). Mice treated with h‐LEH had significantly attenuated inflammatory cytokines, increased surface perfusion, and increased Ki67 expression on day 7 in accordance with the ulcer size reduction, while there was no significant difference in chemokines, histological granulation, epithelial thickness, and granulocyte infiltration detected by immunohistochemical staining in the ulcer between the treatment groups. The results suggest that h‐LEH (2 mL/kg) early after wounding may accelerate skin wound healing in dB/dB mice to levels equivalent to wild‐type mice probably via mechanism(s) involving reduced hypoxia, increased surface perfusion, suppressed inflammation, accelerated in situ cell proliferation and protein synthesis.     

Human pregnancy and generation of anti‐angiotensin receptor and anti‐perlecan antibodies

Non‐HLA antibodies against the angiotensin II type 1 receptor (AT₁R) and the C‐terminal fragment of perlecan (i.e., LG3) are associated with the development of renal allograft rejection. It is currently unknown how humans develop anti‐AT₁R or anti‐LG3 antibodies. The aim of this study was to investigate whether pregnancy—as a model of sensitization to polymorphic proteins—induces anti‐AT₁R and/or anti‐LG3 antibodies. We included 104 samples from women obtained after physiologic full‐term pregnancy and 80 samples from healthy nonsensitized controls (40 women and 40 men). Both anti‐AT₁R and anti‐LG3 antibody levels were lower in pregnancy samples than in controls (both P < 0.05). By multivariate analysis, male gender was an independent predictor for high anti‐AT₁R antibody levels (OR 3.66, P = 0.04) and pregnancy was predictive for low anti‐LG3 antibody levels (OR 6.53, P = 0.0001). There was no correlation of anti‐AT₁R with anti‐LG3 antibody levels, either in the pregnancy or in the control samples (r² ≤ 0.03, P ≥ 0.26). In conclusion, physiologic full‐term pregnancy does not induce anti‐AT₁R or anti‐LG3 antibodies and may even lower their levels. Therefore, anti‐AT₁R and anti‐LG3 antibodies are likely not caused by allosensitization. The lack of correlation of anti‐AT₁R with anti‐LG3 antibodies suggests different mechanisms of generation, which remain to be elucidated.     

Effect of metabolic and antioxidant supplementation on sperm parameters in oligo‐astheno‐teratozoospermia,with and without varicocele: A double‐blind placebo‐controlled study

Since sperm require high energy levels to perform their specialised function, it is vital that essential nutrients are available for spermatozoa when they develop, capacitate and acquire motility. However, they are vulnerable to a lack of energy and excess amounts of reactive oxygen species, which can impair sperm function, lead to immotility, acrosomal reaction impairment, DNA fragmentation and cell death. This monocentric, randomised, double‐blind, placebo‐controlled trial investigated the effect of 6 months of supplementation with l ‐carnitine, acetyl‐l ‐carnitine and other micronutrients on sperm quality in 104 subjects with oligo‐ and/or astheno‐ and/or teratozoospermia with or without varicocele. In 94 patients who completed the study, sperm concentration was significantly increased in supplemented patients compared to the placebo (p = .0186). Total sperm count also increased significantly (p = .0117) in the supplemented group as compared to the placebo group. Both, progressive and total motility were higher in supplemented patients (p = .0088 and p = .0120, respectively). Although pregnancy rate was not an endpoint of the study, of the 12 pregnancies that occurred during the follow‐up, 10 were reported in the supplementation group. In general, all these changes were more evident in varicocele patients. In conclusion, supplementation with metabolic and antioxidant compounds could be efficacious when included in strategies to improve fertility.     

Epidemiology of tick‐borne pathogens in the semi‐arid and the arid agro‐ecological zones of Punjab province,Pakistan

Tick‐borne diseases (TBDs) have a large impact on animal health and the livelihood of livestock owners, particularly in developing countries. Although climatic and ecological conditions in Pakistan may favour the transmission of tick‐borne pathogens (TBPs), only a few systematic studies have been carried out on TBPs and the diseases that they cause in this country. To improve our understanding of the distribution of TBPs, 3,807 ticks were collected from ruminants (n = 369) on 108 livestock farms (semi‐arid zone = 36, arid zone = 72) in Punjab Province. After morphological identification ticks were pooled into 405 pools (Hyalomma anatolicum = 300, Rhipicephalus microplus = 89, Hyalomma dromedarii = 9, Rhipicephalus turanicus = 7) based on their species, locality of collection, and the host. DNA from each pool was screened by a Reverse Line Blot (RLB) hybridization assay for the presence of Anaplasma, Ehrlichia, Rickettsia, Babesia, and Theileria species. DNA from at least one TBP was found in 142 (35.1%) pools. Among the positive pools, 91 (64.1%) had a mixed infection with two or more TBPs, whereas 51 (35.9%) pools were infected with a single TBP. The detected pathogens not only included species that were known to be endemic in Pakistan, such as Theileria annulata (6.7%), Theileria orientalis (3.5%), Anaplasma marginale (5.7%), Anaplasma centrale (2.7%), Anaplasma ovis (1.5%), Babesia bigemina (0.7%), and Babesia bovis (0.2%), but also several TBPs that had not been reported to occur in Pakistan before. This included Ehrlichia minasensis (3.2%), an Anaplasma platys‐like organism (1.2%), Babesia occultans (0.2%), and Rickettsia massiliae (0.2%), as well as two previously uncharacterized species: Ehrlichia sp. Multan (18.0%) and Anaplasma sp. (BL099‐6) (2.22%). The pathogenicity of these novel species remains to be examined. This study shows that a much broader spectrum of TBPs is present in Pakistan than previously thought, including several zoonotic pathogens.     

Difference in outcomes after antibody‐mediated rejection between abo‐incompatible and positive cross‐match transplantations

Graft survival seems to be worse in positive cross‐match (HLAi) than in ABO‐incompatible (ABOi) transplantation. However, it is not entirely clear why these differences exist. Sixty‐nine ABOi, 27 HLAi and 10 combined ABOi+HLAi patients were included in this retrospective study, to determine whether the frequency, severity and the outcome of active antibody‐mediated rejection (AMR) were different. Five‐year death‐censored graft survival was better in ABOi than in HLAi and ABOi+HLAi patients (99%, 69% and 64%, respectively, P = 0.0002). Features of AMR were found in 38%, 95% and 100% of ABOi, HLAi and ABOi+HLAi patients that had a biopsy, respectively (P = 0.0001 and P = 0.001). After active AMR, a declining eGFR and graft loss were observed more frequently in HLAi and HLAi+ABOi than in ABOi patients. The poorer prognosis after AMR in HLAi and ABOi+HLAi transplantations was not explained by a higher severity of histological lesions or by a less aggressive treatment. In conclusion, ABOi transplantation offers better results than HLAi transplantation, partly because AMR occurs less frequently but also because outcome after AMR is distinctly better. HLAi and combined ABOi+HLAi transplantations appear to have the same outcome, suggesting there is no synergistic effect between anti‐A/B and anti‐HLA antibodies.     

Adequacy and Complication Rates with 14‐ vs. 16‐gauge Automated Needles in Percutaneous Renal Biopsy of Native Kidneys

In performing percutaneous renal biopsy (PRB) of native kidneys, an increasing use of 16‐gauge automated biopsy needles has been observed. We compare the adequacy and safety of PRBs in adults performed with a 14‐gauge (n = 82) vs. 16‐gauge (n = 55) automated needle using real‐time ultrasound (US) from 1/2010 to 12/2013. Baseline clinical and laboratory data along with outcome data (renal US 1‐hour postbiopsy, biopsy adequacy, and safety) were collected prospectively. There was no difference in age, gender, blood pressure, serum creatinine, or pre‐PRB hemoglobin at baseline for PRBs performed with a 14‐ vs. 16‐gauge needle. The number of glomeruli obtained per biopsy was similar (29 ± 11 vs. 31 ± 14, p = 0.6) and adequate tissue for diagnosis was obtained in 99% and 100% of biopsies. The clinical complication (8.5% vs. 9.1%, p = 1.0), transfusion (7.3% vs. 7.2%, p = 1.0), and embolization (3.7% vs. 1.8%, p = 0.6) rates were not significantly different for 14‐ vs. 16‐gauge needles, but by routine renal US 1‐hour post‐PRB, a perinephric hematoma was demonstrated more often in biopsies done with the 14‐gauge needle (39% vs. 22%, P 0.04). Thus, while the success of PRB of native kidneys is similar for both needle gauges, the potential for complication may be less using a 16‐gauge automated needle.     

When patients fail UNAIDS’ last 90 ‐ the “failure cascade” beyond 90‐90‐90 in rural Lesotho,Southern Africa: a prospective cohort study

Introduction : HIV‐infected individuals on first‐line antiretroviral therapy (ART) in resource‐limited settings who do not achieve the last “90” (viral suppression) enter a complex care cascade: enhanced adherence counselling (EAC), repetition of viral load (VL) and switch to second‐line ART aiming to achieve resuppression. This study describes the “failure cascade” in patients in Lesotho. Methods : Patients aged ≥16 years on first‐line ART at 10 facilities in rural Lesotho received a first‐time VL in June 2014. Those with VL ≥80 copies/mL were included in a cohort. The care cascade was assessed at four points: attendance of EAC, result of follow‐up VL after EAC, switch to second‐line in case of sustained unsuppressed VL and outcome 18 months after the initial unsuppressed VL. Multivariate logistic regression was used to assess predictors of being retained in care with viral resuppression at follow‐up. Results : Out of 1563 patients who underwent first‐time VL, 138 (8.8%) had unsuppressed VL in June 2014. Out of these, 124 (90%) attended EAC and 116 (84%) had follow‐up VL (4 died, 2 transferred out, 11 lost, 5 switched to second‐line before follow‐up VL). Among the 116 with follow‐up VL, 36 (31%) achieved resuppression. Out of the 80 with sustained unsuppressed VL, 58 were switched to second‐line, the remaining continued first line. At 18 months’ follow‐up in December 2015, out of the initially 138 with unsuppressed VL, 56 (41%) were in care and virally suppressed, 37 (27%) were in care with unsuppressed VL and the remaining 45 (33%) were lost, dead, transferred to another clinic or without documented VL. Achieving viral resuppression after EAC (adjusted odds ratio (aOR): 5.02; 95% confidence interval: 1.14–22.09; p = 0.033) and being switched to second‐line in case of sustained viremia after EAC (aOR: 7.17; 1.90–27.04; p = 0.004) were associated with being retained in care and virally suppressed at 18 months of follow‐up. Age, gender, education, time on ART and level of VL were not associated. Conclusions : In this study in rural Lesotho, outcomes along the “failure cascade” were poor. To improve outcomes in this vulnerable patient group who fails the last “90”, programmes need to focus on timely EAC and switch to second line for cases with continuous viremia despite EAC.     

Seminal plasma miR‐210‐3p is a biomarker for screening dyszoospermia caused by varicocele

To evaluate the value of seminal plasma miR‐210‐3p as a novel and non‐invasive biomarker for screening dyszoospermia caused by varicocele. Semen samples from patients with varicocele and healthy males were collected for semen analysis and quantitative real‐time polymerase chain reaction. Cox univariate and multivariate analysis and receiver operating characteristic curve analysis were used to assess the relationship between the level of seminal plasma miR‐210‐3p and impaired spermatogenic function. Our results showed that the level of seminal plasma miR‐210‐3p in the varicocele patients was 2.18 times that of the control group (p < 0.001), and its expression increased significantly with the severity of varicocele. Compared with preoperative, the expression of seminal plasma miR‐210‐3p declined significantly at 3 months after surgery. Cox univariate and multivariate analysis showed that seminal plasma miR‐210‐3p (p = 0.02), bilateral varicocele (p = 0.04) and the grade 3 varicocele (p = 0.03) were significantly and independently associated with dyszoospermia caused by varicocele. Our results suggest that seminal plasma miR‐210‐3p is a useful clinical biomarker for screening dyszoospermia caused by varicocele, and this is the key to deciding early effective treatment and protecting the fertility of the patients.     

Synthetic hemoglobin‐based oxygen carriers are an acceptable alternative for packed red blood cells in normothermic kidney perfusion

Normothermic machine perfusion presents a novel platform for pretransplant assessment and reconditioning of kidney grafts. Maintaining the metabolic activity of a preserved graft at physiologic levels requires an adequate oxygen supply, typically delivered by crystalloid solutions supplemented with red blood cells. In this study, we explored the feasibility of using a synthetic hemoglobin‐based oxygen carrier (HBOC) in human kidney normothermic perfusion. Fourteen discarded human kidneys were perfused for 6 hours at a mean temperature of 37°C using a pressure‐controlled system. Kidneys were perfused with a perfusion solution supplemented with either HBOC (n = 7) or packed red blood cells (PRBC) (n = 7) to increase oxygen‐carrying capacity. Renal artery resistance, oxygen extraction, metabolic activity, energy stores, and histological features were evaluated. Throughout perfusion, kidneys from both groups exhibited comparable behavior regarding vascular flow (P = .66), oxygen consumption (P = .88), and reconstitution of tissue adenosine triphosphate (P = .057). Lactic acid levels were significantly higher in kidneys perfused with PRBC (P = .007). Histological findings were comparable between groups, and there was no evidence of histological damage caused by the HBOC. This feasibility experiment demonstrates that a HBOC solution can offer a logistically more convenient off‐the‐shelf alternative to PRBC in normothermic machine perfusion of human kidneys.     

High frequency of TTTY2‐like gene‐related deletions in patients with idiopathic oligozoospermia and azoospermia

Genes located on Y chromosome and expressed in testis are likely to be involved in spermatogenesis. TTTY2 is a Y‐linked multicopy gene family of unknown function that includes TTTY2L2A and TTTY2L12A at Yq11 and Yp11 loci respectively. Using PCR amplification, we screened for TTTY2L2A‐ and TTTY2L12A‐associated deletions, in 94 Greek men with fertility problems. Patients were divided into three groups as following: group A (n = 28) included men with idiopathic moderate oligozoospermia, group B (n = 34) with idiopathic severe oligozoospermia and azoospermia, and group C (n = 32) with oligo‐ and azoospermia of various known etiologies. No deletions were detected in group C patients and 50 fertile controls. However, two patients from group A had deletions in TTTY2L2A (7.1%) and six in TTTY2L12A (21.4%), whereas from group B, four patients had deletions in TTTY2L2A (11.8%) and 10 in TTTY2L12A (29.4%). In addition, five patients from both groups A and B (8%) appeared to have deletions in both studied TTTY2 genes, although these are located very far apart. These results indicate that the TTTY2 gene family may play a significant role in spermatogenesis and suggest a possible mechanism of nonhomologous recombinational events that may cause genomic instability and ultimately lead to male infertility.     

Equine Herpesvirus‐1 Myeloencephalopathy,an Emerging Threat of Working Equids in Ethiopia

Although equine herpesvirus myeloencephalopathy (EHM) is a sporadic and relatively uncommon manifestation of equine herpesvirus‐1 (EHV‐1), it has the potential for causing devastating outbreaks in horses. Up till now, there were no reported EHM outbreaks in donkeys and mules. This study describes the isolation and molecular characterization of EHV‐1 from clinically EHM‐affected horses (n = 6), mules (n = 3) and donkeys (n = 82) in Ethiopia during outbreaks from May 2011 to December 2013. The incidence of EHM cases was higher from April to mid‐June. EHM in donkeys was more severe and death without clinical signs of paralysis, and recumbency was frequently observed. The main age of affected equines ranged from 7 to 10 years (n = 51; 56.0%), and females (n = 58; 63.7%) were more affected than males. The incidence of neuropathogenic (D₇₅₂) and non‐neuropathogenic (N₇₅₂) variants of EHV‐1 from EHM‐affected equines in Ethiopia was assessed by sequencing the DNA polymerase gene (ORF30) of the EHV‐1 isolates. The results indicated that from the total of 91 clinically affected equines, 90 (98.9%) of them had an ORF30 D₇₅₂ genotype. An ORF30 N₇₅₂ variant was only found in one donkey. Analysis of ORF68 as grouping marker for geographical differences showed that the Ethiopian EHV‐1 isolates belong to geographical group 4. Due to the fatal nature of EHV‐1 in donkeys, it would be interesting to examine the pathogenesis of EHM in this species. At present, there is no vaccine available in Ethiopia, and therefore, outbreaks of EHV‐1 should be controlled by proper management adaptations. In addition, it is important to test the efficacy of the commercial vaccines not only in horses, but also in donkeys and mules.     

Modified donor lymphocyte infusion‐associated acute graft‐versus‐host disease after haploidentical T‐cell‐replete hematopoietic stem cell transplantation: incidence and risk factors

We performed a study to investigate the profile of donor lymphocyte infusion (DLI)‐associated acute graft‐versus‐host disease (GVHD) in haploidentical T‐cell‐replete hematopoietic stem cell transplantation (HSCT). A total of 124 patients receiving modified DLI after haploidentical T‐cell‐replete HSCT were enrolled. The cumulative incidence of DLI‐associated acute GVHD was 53.2% for grades II–IV and 28.4% for grades III–IV. The duration of GVHD prophylaxis after DLI was the only risk factor for DLI‐associated grades III–IV acute GVHD (p < 0.05). The cumulative incidence of grades III–IV acute GVHD in patients with prophylaxis more than six, four to six, two to four, and <2 wk were 9.3%, 14.4%, 31.6%, and 49.5%, respectively (p = 0.018). Furthermore, DLI‐associated grades III–IV acute GVHD was the only risk factor for overall survival (p = 0.038, OR   = 2.869) and transplant‐related mortality (p = 0.018, OR = 3.296) but not a risk factor for relapse after DLI (p = 0.840). This study confirms for the first time that the duration of GVHD prophylaxis after DLI is the only risk factor for the development of grades III–IV acute GVHD. Donor lymphocyte infusion with prophylaxis more than six wk was associated with a lower incidence of grades III–IV acute GVHD.     

Ten‐year outcomes in a randomized phase II study of kidney transplant recipients administered belatacept 4‐weekly or 8‐weekly

In the phase II IM103‐100 study, kidney transplant recipients were first randomized to belatacept more‐intensive‐based (n = 74), belatacept less‐intensive‐based (n = 71), or cyclosporine‐based (n = 73) immunosuppression. At 3‐6 months posttransplant, belatacept‐treated patients were re‐randomized to receive belatacept every 4 weeks (4‐weekly, n = 62) or every 8 weeks (8‐weekly, n = 60). Patients initially randomized to cyclosporine continued to receive cyclosporine‐based immunosuppression. Cumulative rates of biopsy‐proven acute rejection (BPAR) from first randomization to year 10 were 22.8%, 37.0%, and 25.8% for belatacept more‐intensive, belatacept less‐intensive, and cyclosporine, respectively (belatacept more‐intensive vs cyclosporine: hazard ratio [HR] = 0.95; 95% confidence interval [CI] 0.47‐1.92; P = .89; belatacept less‐intensive vs cyclosporine: HR = 1.61; 95% CI 0.85‐3.05; P = .15). Cumulative BPAR rates from second randomization to year 10 for belatacept 4‐weekly, belatacept 8‐weekly, and cyclosporine were 11.1%, 21.9%, and 13.9%, respectively (belatacept 4‐weekly vs cyclosporine: HR = 1.06, 95% CI 0.35‐3.17, P = .92; belatacept 8‐weekly vs cyclosporine: HR = 2.00, 95% CI 0.75‐5.35, P = .17). Renal function trends were estimated using a repeated‐measures model. Estimated mean GFR values at year 10 for belatacept 4‐weekly, belatacept 8‐weekly, and cyclosporine were 67.0, 68.7, and 42.7 mL/min per 1.73 m², respectively (P<.001 for overall treatment effect). Although not statistically significant, rates of BPAR were 2‐fold higher in patients administered belatacept every 8 weeks vs every 4 weeks.     

Performance of existing risk scores around heart transplantation: validation study in a 4‐year cohort

Several risk scores exist to help identify best candidate recipients for heart transplantation (HTx). This study describes the performance of five heart failure risk scores and two post‐HTx mortality risk scores in a French single‐centre cohort. All patients listed for HTx through a 4‐year period were included. Waiting‐list risk scores [Heart Failure Survival Score (HFSS), Seattle Heart Failure Model (SHFM), Meta‐Analysis Global Group in Chronic Heart Failure (MAGGIC), Organized Program to Initiate Lifesaving Treatment in Hospitalized Patients with Heart Failure (OPTIMIZE‐HF) and Get With The Guidelines‐Heart Failure (GWTG‐HF)] and post‐HTx scores Index for Mortality Prediction After Cardiac Transplantation (IMPACT and CARRS) were computed. Main outcomes were 1‐year mortality on waiting list and after HTx. Performance was assessed using receiver operator characteristic (ROC), calibration and goodness‐of‐fit analyses. The cohort included 414 patients. Waiting‐list mortality was 14.0%, and post‐HTx mortality was 16.3% at 1‐year follow‐up. Heart failure risk scores had adequate discrimination regarding waiting‐list mortality (ROC AUC for HFSS = 0.68, SHFM = 0.74, OPTIMIZE‐HF = 0.72, MAGGIC = 0.70 and GWTG = 0.77; all P‐values <0.05). On the contrary, post‐HTx risk scores did not discriminate post‐HTx mortality (AUC for IMPACT = 0.58, and CARRS = 0.48, both P‐values >0.50). Subgroup analysis on patients undergoing HTx after ventricular assistance device (VAD) implantation (i.e. bridge‐to‐transplantation) (n = 36) showed an IMPACT AUC = 0.72 (P < 0.001). In this single‐centre cohort, existing heart failure risk scores were adequate to predict waiting‐list mortality. Post‐HTx mortality risk scores were not, except in the VAD subgroup.