Search

To provide fresh and highly nutritive food, automated greenhouses and smart farming systems had proved to be helpful for growing world population. The smart greenhouse house monitoring system not only helpful in exploiting the production but also helpful to bridge up the quality of the produce. The current study was designed to explore the potential use of a smart greenhouse monitoring system using Raspberry-Pi microcontroller. The aim of the study was to create a smart automation system to control the microclimate of greenhouse. Two varieties of tomato (Roma and cherry tomato) were used both in smart greenhouse system as well as in conventional greenhouse system to compare the agronomic and quality parameters. Temperature and humidity were set according to the production technology of tomato using automation system. Proper fertilization and irrigation requirements were considered equal in all aspects in smart greenhouse and conventional greenhouse. The smart greenhouse monitoring system worked better in maintaining the microclimate inside the greenhouse with a difference of about 5-6 ℃ temperature and 20-30% humidity higher than the conventional greenhouse. The results predicted a progressive increase in agronomic parameter with a difference of 10-15% in plant height, number of leaves, number of fruits and weight of fruit as compared with growth parameters in conventional greenhouse. Similarly the quality parameters were effective with maximum size of fruit in Roma variety that was 75 mm as compared to fruit size 65 mm in conventional farming. The over average yield of tomato per plant (5.5 kg/plant) was also recorded in smart greenhouse that was significantly increased as compared with conventional greenhouse. The results predicted that the yield of tomato was positively affected using smart greenhouse monitoring system and consequently, the smart technologies could be used for the potential crop production and monitoring of cultivation activities.

To bridge up the quality gaps and excel in the production of eggplant there is a need to switch the conventional production system to smart greenhouse house monitoring system. The study was designed to evaluate the growth parameters using smart greenhouse monitoring system designed with Raspberry-Pi microprocessor and sensors to control the microclimate (temperature and humidity) of a prototype greenhouse. A set of experiment was designed, one in a smart greenhouse system and the 2nd in the conventional greenhouse system to evaluate the growth parameters of eggplant. In a microclimate of smart greenhouse the temperature and humidity was set controlled using two sensors and automation system to control the mist spray and heating/cooling system inside the prototype greenhouse. Standard production technology was adopted in both the experiments and the experiment was replicated thrice. It was observed that the smart greenhouse monitoring system maintained the microclimate inside the prototype greenhouse ultimately effect the growth and yield parameters. The results predicted a progressive increase in growth parameter upto 10-20% in plant height, flowers per plant, fruits per plant and weight of fruit in automated greenhouse using microcontroller. Similarly quality of fruit was significantly affected as maximum fruit diameter 35 mm and fruit firmness 6.9 libre was recorded that was significantly higher than the fruit in the conventional greenhouse. The over average yield of eggplant per plant was recorded 5.6 kg/plant in automated greenhouse while 4 kg/plant fruit yield was recorded in conventional greenhouse that was significantly higher. The findings of current study predicted that the overall yield of eggplant was significantly higher and the growth and quality parameters were also higher in automated greenhouse using microcontroller and can be used for the potential eggplant production in urban agriculture.

The following article outlines the essential clauses within agreements about the funding of German Universities or German clinical trial sites in order for them to conduct a non-commercial clinical trial on their own, a so-called Investigator-Initiated-Trials or Investigator-Sponsored-Trials. The authors explain the basic legal principles and clauses for such an agreement and clarify certain German Law specialities, which any funder should be aware of, if they were to fund an Investigator-Initiated-Trial in Germany. It becomes clear, that it is very important for the funding pharmaceutical company or foundation, not to be confused with the regulatory sponsor of the given clinical trial. Unclear wording in the funding agreement could lead to the actual transfer of a sponsor’s responsibilities from the University or clinical trial site to the funding pharmaceutical company or foundation, with all legal and monetary risks. In order to avoid unwanted penalties and costs, it is imperative for the funding entity to draft the essential clauses carefully. This article aims to help with that.

Globally, optimized doses of exogenously applied growth regulators hold the potential to sustainably boost the growth and productivity of leguminous crops, including green gram. A field investigation was undertaken at the Agronomy Farm of the University of Agriculture Faisalabad, Pakistan in 2021–2022 to determine the highest-performing doses of foliar-applied salicylic acid (S1 = 0 and S2 = 75 ppm) and gibberellic acid (G1 = 0, G2 = 30, G3 = 60, G4 = 90 and G5 = 120 ppm) for green gram (cv. NIAB-MUNG 2011) sown under irrigated conditions in a semiarid climate. The response variables included physiological growth traits (CGR and net assimilation rate (NAR)), yield attributes (plant height (PH), PBs and the number of pods per plant−1 (NP), pod length (PL) and SW, grain (GY) and biological yields (BY), the biosynthesis of pigments (chlorophyll a, chlorophyll b and total chlorophyll along with carotenoids) and protein (P) contents. The results revealed that S2G5 remained unmatched in that it exhibited the highest crop growth rate, while it remained on par with S2G4 and S2G3 in terms of its net assimilation rate. Additionally, S2G5 maximized plant height, the number of pod-bearing branches and pods per plant, pod length, seed number per pod−1 and 1000-seed weight, which led to the highest grain yield and biological yield (104% and 69% greater than those of the control, respectively). Moreover, the same treatment combination also surpassed the rest of the treatments because it recorded the largest amounts of chlorophyll and carotenoid contents, and the P content was increased to 24% greater than that observed for the control treatment. Thus, the exogenous application of salicylic acid (75 ppm) and gibberellic acid (120 ppm) might be recommended to green gram growers to sustainably increase the plant’s yield and nutritional value, and these findings may serve as a baseline for conducting more studies to test higher doses of these growth regulators.

The possibilities of structuring clinical trial agreement are far reaching. Especially regarding the question, who the contract partner will be, an initial, also liability-related, positioning is possible; by either including or not including the respective examining physician in the agreement. The agreement has in large parts a clarifying function regarding the contractual obligations of both parties, as the AMG, the ICH-GCP guideline and various european regulations and directives stipulate a wide range of requirements for the performance of clinical trials. Inclusion in the agreement allows for a clear division of responsibilities and liability in the event of non-compliance with the requirements. It is also necessary that the sponsor is obligated to carry out quality assurance measures and the modalities of their implementation, including the remuneration of the time spent for this purpose. A central question is also, who is entitled to the results of the clinical trial or to inventions based on them, whereby the ArbnErfG also must be taken into account. Whether there are at least rights of use for non-commercial purposes for the other party, should also be regulated. Also relevant is the agreement of confidentiality agreements and the answer to the question, when and to which extent publications on the results of the tests in question are permissible, in order to ensure an appropriate balance of the parties' interests. In terms of data protection law, special attention must be paid to the question which party is the controller within the meaning of the GDPR - depending on the answer to this question, complicated and extensive regulations must be made. In addition to the regulation of the remuneration and its payment modalities, regulations regarding the liability, at least within the legal ramifications are possible and necessary. Conclusively – as always - general cancellation rights and other possibilities of terminating the agreement can be added. The preceding explanations show how complex and comprehensive the aspects to be taken into account are. Numerous possibilities have been highlighted above, in order to provide an overview and to simplify the drafting, in hope that the contribution may aid as an orientation.

The exchange of information and materials within the framework of research projects can be managed in different ways. In most cases a so-called “Material Transfer Agreement” (MTA) is necessary. What looks like a simple ascertainment of rights and duties of the contracting parties, is clearly more fraught with risks than it looks at first sight. An individual arrangement and initial negotiation are necessary to prevent legal risks and not to burden the cooperation between the contracting parties and scientists with legal conflicts and uncertainties. Of course, confidential clauses shouldn’t be missing. While the precise regulations of the scope of work serves the concretion of the subject matter of the contract, clauses for the protection of property and IP rights regulate the handling with the research and possible publications regarding the use of the provided materials and their specific ownerships. The following will deal with the areas of application, the requirements and the regulatory content of this important type of contract.

Dopamine β-hydroxylase (DBH) is a copper-containing enzyme that has an important role in maintaining the cellular homeostasis between the two neurotransmitters, dopamine (DA) and nor-adrenaline (NA). DBH functional polymorphisms are associated with multiple neuro-psychiatric conditions and are found to alter the DBH protein levels in serum, affecting DBH enzymatic activity. The current study was conducted to determine the genetic association of DBH functional polymorphism rs1611115 and its effect on DBH levels in serum in major depressive disorder (MDD), bipolar disorder (BPD) and schizophrenia (SHZ) in the Pakistani population. In total n = 1097 subjects including MDD (n = 427), BPD (n = 204), SHZ (n = 134) and healthy controls (n = 332), were screened for rs1611115 by polymerase chain reaction-restriction fragment length polymorphism. Univariate logistic regression was applied and results were adjusted for age and sex (multivariant analysis). The DBH levels in serum were determined through enzyme-linked immunosorbent assay (ELISA) and the Mann Whitney U test was applied. The results showed a significant association of minor allele (-1021C > T) with a higher risk of developing BPD and SHZ in both univariable and multivariable analyses. Moreover, the overall total serum concentration of DBH was comparatively raised in MDD, however, in cross-comparison DBH serum levels were found markedly higher in CC homozygotes compared to TT homozygotes within the BPD group. Thus the present study suggested a significant association of DBH rs1611115 with BPD and SHZ and also the effect of rs1611115 on DBH serum levels in MDD and BPD, for the first time in the Pakistani population.

Tomato (Lycopersicon esculentum L.) is the second most cultivated vegetable crop. World production of tomato is about 177 million metric tons from 4.78 million ha, and Pakistan contributed an annual production of 2.9 million metric tons from 0.15 million ha (FAO, 2021). The bacterial speck of tomato caused by Pseudomonas syringae pv. tomato has devastated tomato cultivars in terms of their production and quality in Poonch, AJK, Pakistan. In a systemic field survey of tomato growing areas of Poonch division including Rawalakot, Bagh and Hajira, 23 fields were selected randomly with an approximately 10km apart from each other in the temperate climate ranging temperature from 18-22 ℃ with humidity more than 70%. It was observed small black spots with yellow haloes on tomato leaves and on fruits small, dark slightly raised lesions were observed (Mensi et al., 2018). On the basis of symptomological parameters average disease incidence of 42% was observed. Twelve isolates were recovered from the collected leaf, twig and fruit samples on King’s B medium while incubation temperature was 27±2 ℃. All the isolates were gram found gram negative and produced levan when grown on sucrose supplemented medium. Hypersensitive reaction of all the isolates was positive on tobacco leaves and were non-fluorescent under UV light. Pathogenicity test was performed on tomato seedlings inoculated with bacterial suspension having concentration of 108 CFU/ml with five replications and for control treatment sterile distilled water was used. Small dark brown to black spots similar to the symptoms found in the field were observed while no symptoms appeared in the control thus confirming the Koch’s postulates (Valenzuela et al., 2022). The amplification of 16s rRNA and gyrB gene of two isolates was done using 16s rRNA universal primer and an housekeeping gene primer respectively by polymerase chain reaction as described by Karlson et al. (1993) and Yamamoto et al. (2000). The amplicones of 16s rRNA gene were sequenced (GenBank Accession Nos. MK591077 and MK591130) and found 99% identical to the 16s rRNAgene sequences from Pseudomonas syringae pv. tomato. Pathogen was also confirmed by amplification of specific gene sequence gyrB gene (GenBank Accession Nos. MK608712 and MK608713) and successfully amplified 890bp gene sequence confirming the 99% identity upon BLAST. To best of our knowledge it is the first report of Pseudomonas syringae pv. tomato in AJK, Pakistan and was previously reported in Chile (Valenzuela et al., 2022), Tunisia (Mensi et al., 2018), California (Arredondo and Davis, 2000) and in several other countries in the world. The results of the current study will devise further epidemiological studies and quarantine measures to limit the outbreak of this disease on tomato.

Globally, over-reliance on major food crops (wheat, rice and maize) has led to food basket’s shrinking, while climate change, environmental pollution and deteriorating soil fertility demand the cultivation of less exhaustive but nutritious grasses. Unlike neglected grasses (grass species restricted to their centres of origin and only grown at the subsistence level), many underutilized grasses (grass species whose yield or usability potential remains unrealized) are resistant and resilient to abiotic stresses and have multiple uses including food (Coix lacryma-jobi), feed (Eragrostis amabilis and Cynodon dactylon), esthetic value (Miscanthus sinensis and Imperata cylindrica), renewable energy production (Spartina pectinata and Andropogon gerardii Vitman) and contribution to ecosystem services (Saccharum spontaneum). Lack of agricultural market globalization, urbanization and prevalence of large commercial enterprises that favor major grasses trade, improved communication means that promoted specialization in favor of established crops, scant planting material of underutilized grasses and fewer research on their production technology and products development are the prime challenges posed to underutilized grasses promotion. Integration of agronomic research with novel plant protection measures and plant breeding and molecular genetics approaches for developing biotic and abiotic stresses tolerant cultivars along with the development of commercially attractive food products hold the future key for promoting underutilized grasses for supplanting food security and sustainably multiplying economic outcomes.

The dopaminergic pathways control neural signals that modulate mood and behaviour along and have a vital role in the aetiology of major depression (MDD), schizophrenia (SHZ) and bipolar disorder (BD). Genome-wide association studies (GWAS) have reported several dopaminergic pathway’s and other genetic loci’s association with these disorders, therefore, the present study was conducted to analyse the GWAS and candidate gene loci of the dopaminergic and cognitive system genes in MDD, SHZ, and BD, in the Pakistani population. A total of 1237 subjects [MDD n = 479; BD n = 222; SHZ n = 146; and controls n = 390], were screened for eleven genetic variants through polymerase chain reaction (PCR) techniques. Univariant followed by multivariant logistic regression analysis were applied to determine the genetic association. Significant risk associations were observed for rs4532 and rs1799732 with MDD; and rs1006737 and rs2238056 with BD. However, after applying multiple test corrections rs4532 and rs1799732 association did not remain significant for MDD. Moreover, a protective association was found for three variants DRD4-120bp, rs10033951 and rs2388334 in the current cohort. Thus, in conclusion, the current study revealed the risk association of rs1006737 and rs2238056 with BD and the protective effect of DRD4-120bp in MDD and BD, of rs2388334 in BD and of rs10033951 in MDD, BD, and SHZ in the current Pakistani cohort.

Objectives The acute respiratory distress syndrome (ARDS) secondary to viral pneumonitis is one of the main causes of high mortality in patients with COVID-19 (novel coronavirus disease 2019). We systematically reviewed mortality in COVID-19 patients with ARDS and the potential role of systemic corticosteroids in COVID-19 patients. Methods Electronic databases and country-specific healthcare databases were searched to identify relevant studies/reports. The quality assessment of individual studies was conducted using the Newcastle–Ottawa Scale. Country-specific proportion of individuals with COVID-19 who developed ARDS and reported death were combined in a random-effect meta-analysis to give a pooled mortality estimate of ARDS. Results The overall pooled mortality estimate among 10,815 ARDS cases in COVID-19 patients was 39% (95% CI: 23–56%). The pooled mortality estimate for China was 69% (95% CI: 67–72%). In Europe, the highest mortality estimate among COVID-19 patients with ARDS was reported in Poland (73%; 95% CI: 58–86%) while Germany had the lowest mortality estimate (13%; 95% CI: 2–29%) among COVID-19 patients with ARDS. The median crude mortality rate of COVID-19 patients with reported corticosteroid use was 28.0% (lower quartile: 13.9%; upper quartile: 53.6%). Conclusions The high mortality in COVID-19 associated ARDS necessitates a prompt and aggressive treatment strategy which includes corticosteroids. Most of the studies included no information on the dosing regimen of corticosteroid therapy, however, low-dose corticosteroid therapy or pulse corticosteroid therapy appears to have a beneficial role in the management of severely ill COVID-19 patients.

Sugarcane crop constitutes one of the most vital sources of sugar and bioenergy globally; however, higher level of polyploidy makes its genome editing an intricate task. Recently, genome editing has become easier with CRISPR/Cas9 system that uses Cas9 to target sequence-specific regions and introduce double-strand breaks into the target region. This technique has been successfully employed to develop new varieties of sugarcane having desired phenotypic and physiological traits. Several genes can be fused with the CRISPR/Cas9 system leading to successful metabolic engineering and biological improvement for ensuring sustainable enhancement in sugarcane production. This simple RNA-guided genome editing technique has become a revolutionary tool and innovative application in biology that might be effectively employed for inducing specified genomic modifications in plant tissues. This review attempts to synthesize fundamentals of genome editing techniques with an ultimate aim to find out opportunities and challenges of sugarcane genome editing. It also tends to summarize the advances and achievements of gene editing via CRISPR-based genome editing technique for numerous field crops. Lastly, the enormous potential of CRISPR/Cas9 for gene editing in sugarcane, major challenges and future perspectives have been objectively elaborated.

An inherent challenge to clinical trials that aim to test the efficacy of experimental therapeutics for patients with amyotrophic lateral sclerosis (ALS) is the relative rarity of the disease. A promising solution to this problem is a multi-center approach that ideally includes sites distributed across a broad geographic area. In support of such an approach, the European E-RARE program and the United States National Institutes of Health (NIH) partnered to support the investigator-initiated ROCK-ALS trial (Eudra-CT-Nr.: 2017-003676-31, NCT03792490) as a multi-national collaboration between centers in Europe and North America that is led by a European investigator. During the set-up of this international trial, however, a number of unanticipated legal, administrative, and financial complexities emerged that required significant adaptation of the proposed trial scheme. Here, we report our experience navigating these obstacles and describe the potential solutions that we explored. Our experience may inform future efforts to implement multi-national investigator-initiated trials that involve both European and United States centers.

For ensuring food security and imparting sustainability to modern commercial-oriented and highly intensive temperate farming systems, organic wastes from poultry and dairy industries constitute biologically viable strategy to improve crops productivity under changing climate. A field trial was undertaken to appraise the impact of broiler litter (BL = 5 tons ha−1), farm yard slurry (FYS = 10 tons ha−1), and chemical fertilizers including di-ammonium phosphate (DAP = 60 kg ha−1) and single super phosphate (SSP = 60 kg ha−1) applied solely and in conjunction with each other, along with a control treatment (NM). The synergistic fertilization regime encompassing BL+DAP triggered the vegetative growth of soybean as indicated by taller plants having thicker stems and higher leaf area per plant compared to NM. In addition, this fertilization management system improved reproductive yield attributes including pods number and 100-seeds weight which maximized the seed yield, harvest index, seed oil content, and biological yield by 66%, 5%, 31%, and 23% respectively than NM. Moreover, this fertilizers combination was followed by SSP + BL, while BL performed better than FYS and DAP remained superior to SSP. Furthermore, the correlation analyses indicated moderately stronger direct association of seed yield with vegetative growth traits and highly stronger linear relationship with reproductive yield attributes. Thus, co-application of broiler litter (5 tons ha−1) with reduced doses of DAP (60 kg ha−1) might be recommended to temperate soybean growers having access to poultry wastes.

Grapes ( Vitis vinifera ) are the important fruit crop in Pakistan, mostly cultivated for edible purpose. In September 2016, unusual fruit rot symptoms were observed 3-5 days after harvesting on grapes cv . Kishmishi in post-harvest packing houses in Jehlum district (32°56'22.3"N 73°43'31.4"E) of Punjab province. To determine the disease incidence, a total of 10 boxes of grapes from 5 different locations were selected randomly. Each box contained average 12 bunches and 30 bunches out of 120 inspected bunches displayed typical symptoms of the disease. The initial Symptoms were small, round, water-soaked lesions that rapidly developed into soft, white to light pink mycelium near the centre of infected fruits (Figure 1). A total of 186 symptomatic berries were surface sterilized with 1% sodium hypochlorite, rinsed three times with sterile distilled water and dried by placing on filter paper for 45 sec. Sterilized tissues (approximately 4 mm 3 ) were excised and incubated on potato dextrose agar (PDA) medium at 25 ± 4°C. One week after incubation, colonies with abundant aerial mycelium were initially white, cottony and turned to violet and dark purple with age (Figure 2). A total of 25 isolates were examined morphologically. Macroconidia were slender, thin-walled, 3 to 5 septate, curved apical cell, with 20.9 to 45.2 × 3.2 to 7.1 μm and Microconidia were thin-walled, aseptate, club-shaped with 4.5 to 11.2 × 2.3 to 4.1 μm (Figure 3). These characteristics best fit for the description of Fusarium proliferatum (Leslie and Summerell, 2006). Portions of the internal transcribed spacer (ITS) region were sequenced (White et al. , 1990). Sequences of two isolates Fus 07 and Fus 09 (GenBank Accessions; MH444366 and MH464139) showed 100% identity to the corresponding gene sequences of Fusarium proliferatum (GenBank Accessions; MH368119, MF033172 and KU939071) (Figure 4). Pathogenicity test was performed by inoculation with 50-μl conidial suspension (1 × 10 6 conidia/ml) of two isolates onto three non-wounded and four wounded asymptomatic grapes berries. Sterile distilled water was used for a negative control (Figure 5). The experiment was conducted twice and berries were incubated at 25 ± 2°C in sterile moisture chambers (Ghuffar et al. , 2018). White to light pink mycelium in appearance with the original symptoms were observed on both wounded and non-wounded inoculated berries after 3 days, whereas no symptoms were observed on the negative control. The morphology of the fungus that was re-isolated from each of the inoculated berries was identical to that of the original cultures. Fusarium proliferatum, one of the destructive species, causes diseases like foot-rot of corn (Farr et al. , 1990), root rot of soybean (Diaz Arias et al. , 2011), bakanae of rice (Zainudin et al. , 2008), wilt of date palm (Khudhair et al. , 2014), tomato wilt (Chehri, 2016) and tomato fruit rot (Murad et al. , 2016). To our knowledge, this is the first report of Fusarium proliferatum causing fruit rot of grapes in Pakistan, where the disease poses a significant threat to the sustainability of this major fruit crop.

Under the changing climate, fertilization regimes and weed infestation management in aromatic direct-seeded fine rice (DSR) remain vital for curbing environmental hazards and ensuring food security. A multi-year field study was undertaken to appraise the influence of fertilization techniques and weed-free periods on weed dynamics, nutrient uptake and paddy yield in a semi-arid environment. Treatments included two fertilization methods (broadcasting and side placement) and five weed-free durations (20, 30, 40, 50 post-seeding days, DAS) along with a weed-free crop for a whole season. Weed competition for a season-long crop (weedy check) was maintained for each fertilizer application method. Our results revealed that the side placement of fertilizers resulted in a significantly lower weed density and biomass, even under season-long weed competition. The highest paddy yield was recorded for a crop without weeds, while weed-free duration of up to 50 DAS followed it. The uptake of nitrogen (N), phosphorus (P) and potassium (K) for a weed-free duration of up to 50 DAS were only 19%, 9% and 8%, respectively, as compared to the weedy check. The uptake of N, P and K by weeds in the broadcast method was 18%, 30% and 24% higher, compared to side-placed fertilizers. The period of 20–50 DAS remained critical in DSR as far as weed control was concerned. Thus, the side placement of fertilizers and controlling weeds for up to 50 days after rice sowing can be recommended for general adoption in semi-arid agro-ecological conditions.

Impaired neural plasticity may be a core pathophysiological process underlying the symptomatology of schizophrenia. Plasticity-enhancing interventions, including repetitive transcranial magnetic stimulation (rTMS), may improve difficult-to-treat symptoms; however, efficacy in large clinical trials appears limited. The high variability of rTMS-related treatment response may be related to a comparably large variation in the ability to generate plastic neural changes. The aim of the present study was to determine whether negative symptom improvement in schizophrenia patients receiving rTMS to the left dorsolateral prefrontal cortex (DLPFC) was related to rTMS-related brain volume changes. A total of 73 schizophrenia patients with predominant negative symptoms were randomized to an active (n=34) or sham (n=39) 10-Hz rTMS intervention applied 5 days per week for 3 weeks to the left DLPFC. Local brain volume changes measured by deformation-based morphometry were correlated with changes in negative symptom severity using a repeated-measures analysis of covariance design. Volume gains in the left hippocampal, parahippocampal and precuneal cortices predicted negative symptom improvement in the active rTMS group (all r⩽-0.441, all P⩽0.009), but not the sham rTMS group (all r⩽0.211, all P⩾0.198). Further analyses comparing negative symptom responders (⩾20% improvement) and non-responders supported the primary analysis, again only in the active rTMS group (F(9, 207)=2.72, P=0.005, partial η 2=0.106). Heterogeneity in clinical response of negative symptoms in schizophrenia to prefrontal high-frequency rTMS may be related to variability in capacity for structural plasticity, particularly in the left hippocampal region and the precuneus.

The article aims to give an overview of the contractual situation in Germany, which arise in clinical studies. The goal of the authors is to show any potential sponsor of a clinical study, who is interested in cooperation with German partners such as universities or other study sites, what kind of agreements may be expected and to give a brief overview about necessary themes included within the contractual negotiations. The different contractual settings are considered from the perspective each different type of agreement. The authors have chosen amongst all possible types the ones which are the most common in a clinical study, such as CDA, MTA, grant agreement, clinical study agreement, CRO-service agreement etc. The authors show the complexity of the contractual setting of a clinical study and emphasize to pay close attention to the contractual settings. Even though the content of the agreement is mostly universal in nature, in some cases, Germany has some very special rules (e.g. regarding employees’ inventions), which any potential sponsor should keep in mind.

Background: The variability of responses to plasticity-inducing repetitive transcranial magnetic stimulation (rTMS) challenges its successful application in psychiatric care. No objective means currently exists to individually predict the patients' response to rTMS. Methods: We used machine learning to develop and validate such tools using the pre-treatment structural Magnetic Resonance Images (sMRI) of 92 patients with schizophrenia enrolled in the multisite RESIS trial (http://clinicaltrials.gov, NCT00783120): patients were randomized to either active (N = 45) or sham (N = 47) 10-Hz rTMS applied to the left dorsolateral prefrontal cortex 5 days per week for 21 days. The prediction target was nonresponse vs response defined by a >= 20% pre-post Positive and Negative Syndrome Scale (PANSS) negative score reduction. Results: Our models predicted this endpoint with a cross-validated balanced accuracy (BAC) of 85% (nonresponse/response: 79%/90%) in patients receiving active rTMS, but only with 51% (48%/55%) in the sham-treated sample. Leave-site-out cross-validation demonstrated cross-site generalizability of the active rTMS predictor despite smaller training samples (BAC: 71%). The predictive pre-treatment pattern involved gray matter density reductions in prefrontal, insular, medio-temporal, and cerebellar cortices, and increments in parietal and thalamic structures. The low BAC of 58% produced by the active rTMS predictor in sham-treated patients, as well as its poor performance in predicting positive symptom courses supported the therapeutic specificity of this brain pattern. Conclusions: Individual responses to active rTMS in patients with predominant negative schizophrenia may be accurately predicted using structural neuromarkers. Further multisite studies are needed to externally validate the proposed treatment stratifier and develop more personalized and biologically informed rTMS interventions.

An elderly, bed ridden patient with a history of stroke was admitted for management of aspiration pneumonia. Two days after insertion of a nasopharyngeal airway, sudden respiratory distress prompted further investigations which led to the eventual diagnosis and removal of the nasopharyngeal airway that had been aspirated. The device was removed under conscious sedation with fiber optic bronchoscopy. Keywords: Nasopharyngeal airway, Aspiration, Respiratory distress, X-ray

Gray mold caused by Botrytis cinerea is an important disease that attacks fruits, leaves and twigs of peach. Peach is grown on an area of 18,008 ha with an average production of 72,085 tons per year in Pakistan (FAO, 2017). During May 2017, brown spots on 33% of the peach fruits examined were observed in Swat district of KPK province of Pakistan. Infected fruits were incubated at 25±2 °C in a humid chamber resulted in greyish mycelial growth with light brown lesions. Hyphal growths on infected fruits were cultured on PDA media and purified by hyphal tip method. Morphologically whitish grey growth was observed on PDA and later on dark sclerotia were observed after 6-7 days of incubation. Hyphae were found septate with branched hyaline conidiophores having a bunch of ovoid conidia at their tips. Further confirmations were done by amplifying internal transcribed spacer regions (Andrew et al. , 2009) and glyceraldehyde-3-phosphate dehydrogenase (G3PDH) region of the isolates (Li et al. , 2012). Amplicons sequenced from Macrogen Korea were blasted and submitted in NCBI showed that ITS sequences (Accessions MH049690 and MH049691) were 99% identical with already reported (MG878388 and MG654661) sequences and the G3PDH gene sequences (Accessions MH560352 and MH560353) were 99 % identical with already reported (Accessions MG204876) sequences of B. cinerea . Pathogenicity was confirmed on healthy peach fruits disinfected with 50% ethanol, inoculated by placing a plug of about 1cm 2 taken from the edge of actively growing B. cinerea isolate (BTS-16). Fruits were incubated at 25±2 °C in a humid chamber (Abata et al. , 2016). A set of healthy fruits mock-inoculated with a plug of agar medium were used as control. Three days after inoculation, inoculated fruits showed sunken lesions with cottony greyish mycelial growth on their surface. Fungus isolated from these infections was re-confirmed as B. cinerea . Conducive environment for the disease progression in nearby areas can result into a huge loss in peach produce so there is a need to devise management strategies to cope with the pathogen. This is the first report of gray mold disease of peach caused by B. cinerea from Pakistan.

Bell pepper ( Capsicum annuum L.) is one of the extensively cultivated vegetable crop in Punjab, Pakistan. During two years of field surveys, February-November 2016-17, damping off and blight symptoms were observed. Average seedling mortality was recorded as 18.7% while yield loss due to blight was estimated 32 to 41% at mature stages. Maximum blight infection was recorded from the areas frequently flooded with canal irrigation system. At early stages, lesions were noticed on stem portions at soil line level while at crop maturity stages blight symptoms were noted. Leaves were blanched and wilted while fruits were covered with white mold. Masses of sporangia were evident on and inside the infected fruits under humid conditions. A total of twelve isolates were recovered from infected root, stem and fruit portions on rye agar media (Caten and Jinks, 1968) incubated at 25 o C under fluorescent light. Papillated sporangia were averaged 42 ± 2.6 X 27 ± 1.7 μm in size (range 27 - 52 × 23 - 36 μm). Oospores were produced on 20% V8 agar and were spherical 22 ± 1.4 μm in diameter (range 14 to 27 μm) while average pedicels length was recorded as 58 ± 12.5 μm (range 13 to 120 μm). These observations were similar to those described for P. capsici (Cocoa, 1988). DNA was extracted using Cetyl Trimethylammonium Bromide (CTAB) method and the internal transcribed spacer regions (ITS1 and ITS2) were amplified by polymerase chain reaction (White et al. , 1990). The amplicons were purified and sequenced in both directions (GenBank Accession No. MF322868 and MF322869). BLAST analysis revealed these isolates showed 99% identity with ITS sequences of Phytophthora capsici (KM369964 and KU518782). Pathogenicity assay was performed on healthy bell pepper seedlings with five repeats. Soil was flooded with 20ml sporangial suspension (1 x 10 3 sporangia/ml) in pots containing seedlings while 5ml suspension was sprayed until run off on mature plants (Hyder et al. , 2018). A set of uninoculated seedlings was used as control. Pots were kept in dew chamber for 10-20 days at 25±2 o C. Seedling mortality was observed five days after inoculation while at later stage plants develop brown-to-black stem lesions with white mycelial growth on leaves. These symptoms were identical to the P. capsici infections in field. Consistent re-isolations of P. capsici confirm its association with the disease. To our knowledge, this is the first report of Phytophthora blight on bell pepper from Pakistan

Background Repetitive transcranial magnetic stimulation (rTMS) applied to the left frontal lobe is discussed to be a promising add-on treatment for negative symptoms in schizophrenia. The Positive and Negative Syndrome Scale (PANSS) has been used as outcome parameter in several previous rTMS trials, but studies focusing on PANSS factor analyses are lacking. For this purpose, we used the available PANSS data of the ‘rTMS for the Treatment of Negative Symptoms in Schizophrenia’ (RESIS) trial to calculate different literature-based PANSS factors and to re-evaluate the impact of rTMS on negative symptoms in this trial. Methods In an exploratory re-analysis of published data from the RESIS study (Wobrock et al. 2015), we tested the impact of rTMS applied to the left dorsolateral prefrontal cortex on two PANSS factors for negative symptoms in psychotic disorders as well as on a PANSS five-factor consensus model intending to show that active rTMS treatment improves PANSS negative symptom subscores. Results In accordance to the original analysis, all PANSS factors showed an improvement over time in the active and, to a considerable extent, also in the sham rTMS group. However, comparing the data before and directly after the rTMS intervention, the PANSS excitement factor improved in the active rTMS group significantly more than in the sham group, but this finding did not persist if follow-up data were taken into account. These additional analyses extend the previously reported RESIS trial results showing unspecific improvements in the PANSS positive subscale in the active rTMS group. Our PANSS factor-based approach to investigate the impact of prefrontal rTMS on different negative symptom domains confirmed no overall beneficial effect of the active compared to sham rTMS. Discussion This secondary analysis of the RESIS trials has several limitations. First of all, the analysis of the primary endpoint was negative [24] and all subsequent secondary analyses showing a positive effect of the intervention (here: change in PANSS excitement factor) are of limited statistical power and therefore subject to uncertainty. On the other hand, our analyses confirm the negative finding of the original publication extends this finding to a broader negative symptom definition. Moreover, the new analysis provides a possible, but hypothetical explanation for the previously described effect of active rTMS on PANSS positive subscale. Of course, many other PANSS factor models are available and in pharmacological research the Marder factors [23, 35] have particular significance. However, the here used five-factor consensus model [21] includes the Marder factor results and our negative symptom factors overlaps with those factors. Another limitation is that it may be possible that our sham stimulation (coil tilted over one wing at an angle of 45°[24]) may still have been slightly biologically active as discussed elsewhere [24].

To assess the safety of direct trocar insertion (DTI) before creating pneumoperitoneum in laparoscopic procedures.An observational study.Combined Military Hospital (CMH) Kohat, Pakistan from November 2013 to January 2015.All the cases undergoing laparoscopy at the study centre in the above duration were included in the study after approval from the Hospital Ethical Committee and informed written consent. Out of the 200 cases, DTI was successfully used to establish peritoneal access in 190 cases, while open Hassen's technique (OL) was used in ten cases. Body mass index (BMI) of all the patients was calculated before surgery. All the cases were performed under general anaesthesia with adequate relaxation. A 10 mm permanent re-useable sharp metallic trocar was inserted through umbilical scar after lifting the abdominal wall using towel clamps. Studied variables included age, gender, BMI, operations, history of previous surgery, number of attempts for DTI and complications.Mean age of the patients in DTI group was 46.58 ±13.94 years, while 48.70 ±10.08 years in OL group. Female to male ratio in DTI group was 1.43:1; and 2.33:1 in OL group. Increase in BMI had a relation with number of unsuccessful attempts of DTI, so obesity was the main reason of failure of DTI.DTI is a safe and effective method of peritoneal access for laparoscopy with very low failure rate related to BMI and minimal complications.

Background Very few data exist on the management of community-acquired pneumonia (CAP) in patients admitted to hospitals in the Gulf region. The objectives of this study were to describe treatment patterns for CAP in 38 hospitals in five Gulf countries (United Arab Emirates, Kuwait, Bahrain, Oman, and Qatar) and to compare the findings to the most recent Infectious Diseases Society of America (IDSA) / American Thoracic Society (ATS) guidelines. Methods This was a prospective, observational study conducted between January 2009 and February 2011. Adult patients hospitalised (excluding intensive care units) for CAP and subsequently discharged were included. Data were collected retrospectively at hospital discharge, and prospectively during two follow-up visits. Data on medical history, mortality-risk scores, diagnostic criteria, antibiotic treatment, isolated pathogens and clinical and radiographic outcomes were collected. Care practices were compared to the IDSA/ATS guidelines. Results A total of 684 patients were included. The majority (82.9 %) of patients were classified as low risk for mortality (pneumonia severity index II and III). The majority of patients fulfilled criteria for treatment success at discharge, although only 77.6 % presented a normalised leukocyte count. Overall, the management of CAP in Gulf countries is in line with the IDSA/ATS guidelines. This applied to the diagnosis of CAP, to the identification of high-risk CAP patients, to the identification of etiologic agent responsible for CAP and to the type of treatment despite the fact that combinations of antimicrobial agents were not consistent with the guidelines in 10 % of patients. In all patients, information about Gram’s staining was not captured as recommended by the IDSA/ATS and in the majority of patients (>85 %) chest radiography was not systematically performed at the post-discharge follow-up visits. Discussion The management of CAP in the Gulf region is globally in line with current IDSA/ATS guidelines, although rates of pathogen characterisation and post-discharge follow-up need to be improved. Conclusion Compliance with established guidelines should be encouraged in order to improve the management of the disease in this region.

Because of increased safety concerns about synthetic antioxidants, exploitation of cheaper and safer sources of antioxidants based on natural origin is the focus of research nowadays. Rhazya stricta is a medicinally important plant native to South Asia. Extraction of antioxidants was carried out in different solvent systems, i.e., water, 80% methanol, 70% ethanol, and diethyl ether. The methanolic extract exhibited the highest total phenolic content among the extracts; therefore for further studies the methanolic extract was employed. Antioxidant activity measurement in the linoleic acid system, metal chelating activity, reducing power, scavenging effect on 1,1-diphenyl-2-picrylhydrazyl free radical, and superoxide anion radical scavenging activity were taken as the parameters for assessment of antioxidant potential of methanolic extracts. Results were compared with alpha-tocopherol and the synthetic antioxidant butylated hydroxyanisole. The antioxidant potential of methanolic extracts of R. stricta leaves was comparable with previously exploited potent antioxidants and is strongly concentration dependent.

We report the first case of native aortic and mitral valve endocarditis due to Gemella bergeriae from the Middle East in a young patient with rheumatic heart disease. Our case illustrates a fulminant course of infection with G. bergeriae endocarditis that was complicated by embolic stroke, as well as intracerebral and subarachnoid haemorrhage secondary to rupture of a mycotic aneurysm in the right middle cerebral artery. This case highlights the dire, unreported neurological complications of infective endocarditis due to a rare causative organism—G. bergeriae.

Objective. To determine the impact of an intensivist lead tracheostomy care team on the number of successful decannulations in non-critically ill patients and the time required for such decannulations and discharge from the hospital. Methods. Following the introduction of a multidisciplinary tracheostomy care task force, data was collected prospectively from July to December 2009. Matching control data was collected retrospectively from January to June 2009. Chi-Square and Mann-Whitney U- test were used to compare the differences in study variables with an α of 0.05. Results. A total of 44 and 47 patients with percutaneous tracheostomy (PCT) were discharged from the ICU pre- and post-implementation of the task force, respectively. Nine patients in pre- and 24 in post- implementation phase were decannulated, discharged from the hospital, and repatriated to their country of origin (Chi Sq. = 9.21, P = .002). Patients in pre-implementation group had longer hospital stay post ICU discharge compared to patients in post-implementation (58 vs. 34.1 days) group ( P< .001). Conclusion. Structured multidisciplinary tracheostomy care team was associated with significant increase in number of decannulations, hospital discharges, and repatriation post ICU discharge.

Introduction. Retrospective observational data show that ACE-inhibitor therapy delays renal failure and improves life expectancy in Alport patients with proteinuria. The EARLY PRO-TECT Alport trial assesses the safety and efficacy of early therapy onset with ramipril in pediatric Alport patients. Methods and analysis. This double-blind, randomized, placebo-controlled, multicenter phase III trial (NCT01485978; EudraCT-number 2010-024300-10) includes 120 pediatric patients aged 24 months to 18 years with early stages of Alport syndrome (isolated hematuria or microalbuminuria). From March 2012, up to 80 patients will be randomized 1:1 to ramipril or placebo. In the event of disease progression during 3-year treatment, patients are unblinded and ramipril is initiated, if applicable. Approximately 40 patients receive open-label ramipril contributing to the safety database. Primary end-points are “time to progression to next disease level” and “incidence of adverse drug events before disease progression.” Treatment effect estimates from the randomized comparison and Alport registry data will be combined in supportive analyses to maximize evidence. Conclusion. Without this trial, ACE inhibitors may become standard off-label treatment in Alport syndrome without satisfactory evidence base. The results are expected to be of relevance for therapy of all pediatric patients with kidney disease, and the trial protocol might serve as a model for other rare pediatric glomerulopathies.

Background Systemic inflammation can be associated with a redistribution of organ blood flow and a decrease in gastrointestinal perfusion. Regional sympathetic blockade by means of thoracic epidural anesthesia (TEA) has been shown to improve intestinal microcirculation during systemic inflammation. This study tests the hypothesis that during systemic inflammation, TEA attenuates the impairment of gastrointestinal organ perfusion without compromising blood flow to vital organs. Methods Eighteen rats were anesthetized, hemodynamically monitored, and mechanically ventilated with room air. By using fluorescent microspheres, organ perfusion was quantified at baseline, 30 min after the start of epidural infusion of either 2% lidocaine (TEA) or normal saline (control), and after 60 and 120 min of intravenous Escherichia coli lipopolysaccharide infusion in TEA and control animals. Results Blood pressure initially was lower in TEA animals, but it was comparable to controls during endotoxemia. Gastrointestinal organ perfusion significantly decreased after 120 min of endotoxemia in the controls but not in the TEA animals (-23 +/- 27% vs. -6 +/- 26%, mean +/- SD, P < 0.05). Perfusion of the vital organs such as the heart, brain, liver, and kidneys was comparable between controls and TEA after 120 min of endotoxemia. Conclusions TEA attenuates the impairment of gastrointestinal organ perfusion during endotoxemia. Hence, the protective effects of TEA on intestinal microcirculation during endotoxemia may be due to a higher total organ blood flow compared with endotoxemic control animals. Furthermore, in the course of endotoxemia, TEA provides hemodynamic stability and does not compromise blood flow to vital organs.

Objective: We assessed the safety and complications related to percutaneous tracheostomy (PCT) without bronchoscopic guidance in our intensive care unit (ICU). Methods: The prospective data over a period of 24 months were collected for patients who underwent PCT. Major, minor and long-term complications were recorded. The parameters recorded were: age, gender, Glasgow Coma Scale (GCS) score on the day of tracheostomy, acute physiology and chronic health evaluation II (APACHE) score, and predicted mortality based on score on admission and on the day of procedure, number of days on ventilator before and after the procedure, total number of days in the hospital before the final outcome, number of successful decannulations and mortality. The patients were stratified in two groups of survivors and nonsurvivors. Results: A total of 117 patients underwent PCT. Overall mean GCS and APACHE-II scores before PCT were 7 ± 3 and 16 ± 5, respectively. The only significant difference was APACHE-II score and the predicted mortality based on APACHE-II score on the day of PCT, which was higher amongst the nonsurvivors (P = 0.008 and P = 0.006). All 57 (49%) survivors were successfully decannulated with mean post tracheostomy days of 24 ± 15. The major complication observed was three episodes of major bleeding. Only six patients had an episode of desaturation during the procedure and there were three episode of accidental puncturing of endotracheal (ET) tube pressure cuff. During subsequent follow-up in hospital, six patients developed stomal cellulitis. Conclusions: PCT without bronchoscopic guidance can be performed safely by carefully selecting patients and having an experienced team High APACHE score on the day of procedure may lead to poor outcome.

To determine the correlation and diagnostic utility of impaired left atrial emptying fraction, an indicator of diastolic dysfunction, with raised intravascular volume determined by vascular pedicle width on upright postero-anterior chest roentgenogram.Cohort study.Department of Medicine, King Abdulaziz National Guard Hospital, Alhasa, Saudi Arabia from October till December 2002.Thirty-two out of 56 patients with normal systolic function (ejection fraction50%) on echocardiogram were included in the study. Left atrial dimensions were measured at the end of left ventricular diastole (LADed) and systole (LADes) on M-mode tracing. Left atrial emptying fraction (LAEF) was calculated by dividing LADed by LADes. Ratio of peak early diastolic velocity (E/A) to peak late diastolic velocity, isovolemic relaxation time (IVRT) and deceleration time of E-wave (DT) were also recorded on transmitral Doppler signal. Postero-anterior views of chest X-rays done within 48 hours of echocardiogram were studied for the measurement of vascular pedicle width (VPW) and cardiothoracic ratio (CTR). Univariate and multiple regression analysis were performed to identify possible predictors of VPW. Receiver operator curves were constructed to see diagnostic utility of impaired LAEF for raised intravascular volume measured by VPW.Mean age for the patients was 58.56 +/- 17 years. There were 15 females (46.9%) and 17 males (54.1%). LADes, LADed, LAEF and DT were 2.89 +/- 0.63 cm, 3.81 +/- 0.53 cm, 0.75 +/- 0.10 and 202 +/- 49.9 msec respectively. VPW and CTR were 55.81 +/- 10.2 mm and 0.54 +/- 0.08. Significant independent predictors of VPW on univariate regression analysis were entered into multiple regression models, which revealed LAEF and DT as the only predictors of VPW. The final regression equation was VPW = 15.43 + 35.7 (LAEF) + 0.06 (DT), R2 = 0.35, p = 0.0008. LAEF0.75 had a sensitivity of 74% and specificity of 94% for diagnosing raised intravascular volume defined as VPW53 mm.LAEF0.75 is a reliable indicator of intravascular volume expansion in patients with normal systolic function signifying diastolic dysfunction and activation of salt and water retaining mechanisms.

Whole lung lavage is still the most effective treatment for pulmonary alveolar proteinosis. We report a 21-year-old male diagnosed with pulmonary alveolar proteinosis by open lung biopsy and who underwent whole lung lavage with a modified technique. He showed significant improvement in clinical and functional parameters. The technique of intermittent double lung ventilation during lavage procedure keeps the oxygen saturation in acceptable limits in patients at risk for severe hypoxemia and allows the procedure to be completed in a single setting.